CN1898212B

CN1898212B - Analogs of benzoquinone-containing ansamycins for the treatment of cancer

Info

Publication number: CN1898212B
Application number: CN2004800386529A
Authority: CN
Inventors: J·亚当斯; Y·高; A·T·乔治斯埃文格利诺斯; L·格雷尼尔; R·H·朴; J·R·波特; J·L·赖特
Original assignee: Infinity Discovery Inc
Current assignee: Infinity Discovery Inc
Priority date: 2003-12-23
Filing date: 2004-12-23
Publication date: 2012-06-13
Anticipated expiration: 2024-12-23
Also published as: ZA200604863B; CN1898212A

Abstract

The present invention provides analogs of benzoquinone-containing ansamycins and uses thereof for treating and modulating disorders associated with hyperproliferation, such as cancer ( formula (I) and (IV). The present invention provides analogs of benzoquinone-containing ansamycins where the benzoquinone is reduced to a hydroquinone and trapped by reaction with a suitable acid, preferably ones that increase the solubility and air stability of the resulting (17)-ammonium hydroquinone ansamycin analog.

Description

Be used to treat the similar thing of the Ansamycin that comprises benzoquinones of cancer

Related application

The application requires to enjoy following preference: U.S. Provisional Application sequence number 60/532,080, and the applying date is on December 23rd, 2003; U.S. Provisional Application sequence number 60/540,142, the applying date is on January 29th, 2004; U.S. Provisional Application sequence number 60/547,381, the applying date is on February 23rd, 2004; U.S. Provisional Application sequence number 60/561,718, the applying date is on April 12nd, 2004; U.S. Provisional Application sequence number 60/567,565, the applying date is on May 3rd, 2004; U.S. Provisional Application sequence number 60/606,283, the applying date is on September 1st, 2004; U.S. Provisional Application sequence number 60/626,286, the applying date is on November 9th, 2004; U.S. Provisional Application sequence number 60/632,858; The applying date is on December 3rd, 2004; Its specification sheets all is incorporated herein by reference at this.

Background technology

NSC 122750 is a kind of Macrocyclic lactams, and it is the natural Ansamycin family member who comprises benzoquinones.The separation of NSC 122750, preparation and multiple application are described in U.S. Patent number 3,595, in 955.The same with naturally occurring this molecule type of majority member; NSC 122750 generates (Joumal of Antibiotics Vol.23, Page 442 (1970)) as the tunning of Streptomyces hygroscopicus var.geldanus var.nova strain usually.Identified or synthesized the analogue and the verivate of other NSC 122750, they are described in U.S. Patent number 4,261 as the application of antitumour drug, 989 and 5,387,584 and disclosed PCT application WO 00/03737 and WO 03/072794 in.A member in this family of The effect is 17-allyl amino-17-demethoxylation NSC 122750 (" 17-AAG ").NSC 122750 and verivate thereof have shown and have been connected in HSP90 and this protein-active of antagonism.

HSP90 is highly abundant albumen, and it is absolutely necessary for cell viability and has shown dual accompaniment functions (dual chaperone functions) (J.Cell Biol. (2001) 154:267-273, Trends Biochem.Sci. (1999) 24:136-141).It is through interacting with multiple protein behind the native conformation that is changed them by multiple environmental stress such as heat shock; In cellular stress, play a crucial role, guaranteed that enough albumen is overlapping and prevented non-specific set (Pharmacological Rev. (1998) 50:493-513).In addition, the result shows that HSP90 also can work to slowing down the sudden change influence recently, and supposition is through proofreading and correct varient proteinic improper folding (Nature (1998) 396:336-342).In addition; HSP90 also plays important regulating effect under normal physiological conditions; Determining the conformational stability and the sudden change of a large amount of special client's albumen (clientprotein), wherein known about 40 kinds (referring to, Expert.Opin.Biol Ther. (2002) 2 (1): 3-24).This can be divided into three groups in the Asia: uncorrelated proteic set is gone up on steroid hormone acceptor, serine/threonine or Tyrosylprotein kinase and surface, comprises the catalytic subunit of p53 two mutants and Telomere terminal transferase hTERT.All these albumen play regulating effect in cell physiological and biological process.

In large number of biological develops relation, studied the HSP90 antagonist at present, wherein can obtain for a certain state of an illness or disorderly curative effect through suppressing the active one or more aspects of HSP90.Although mainly focus on proliferative disorders such as cancer, other state of an illness is just showing the treatment level that uses the HSP90 antagonist.For example, U.S. Patent application 2003/0216369 discloses the HSP90 suppressor factor to be used to treat virus disorderly.The HSP90 suppressor factor also involves many other application, comprises as antiphlogiston, treatment autoimmunization medicine, treatment apoplexy, local asphyxia, cardiac disorders medicine and be applicable to promoting drug for nerve regeneration (for example referring to, WO 02/09696 (PCT/US01/23640); WO 99/51223 (PCT/US99/07242); USP 6,210,974 B1; And USP 6,174,875).Bibliographical information include but not limited to that fiber (fibrogenetic) disorder that becomes of scleroderma, polymyositis, general lupus, rheumatoid arthritis, chronic inter stitial hepatitis, keloid generation, interstitial nephritis and pulmonary fibrosis can use the HSP90 inhibitor for treating.(Strehlow，WO?02/02123；PCT/US01/20578)。

The nmole of NSC 122750 is renderd a service and the apparent selectivity of kill tumor cell and its main target spot in mammalian cell of being found are HSP90, has excited to develop it and be the interest of antitumour drug.But the utmost point low water solubility of these molecules and the relevant liver toxicity that gives NSC 122750 cause being difficult to develop ratified of the medicine that supplies treatment to use.Especially, NSC 122750 is water-soluble relatively poor, makes it be difficult to carry with the treatment effective dose.

Recently, concentrate on the 17-aminoderivative, particularly 17-AAG of NSC 122750, it is illustrated in keeps the active liver toxicity that reduced simultaneously of HSP90 combination.Referring to U.S. Patent number 4,261,989,5,387,584 and 5,932,566.Be similar to NSC 122750,17-AAG's is water-soluble very limited.This character causes needs to use solubilization carrier, for example Yelkin TTS and DMSO, or Cremophore_ (BASF Aktiengesellschaft) (a kind of ethoxyquin Viscotrol C that gathers); The existence of these carriers causes occurring in some patient's body serious side reaction.

Therefore, still need develop solubleness and better comprise the similar thing of Ansamycin of benzoquinones and concrete grammar and general method, particularly NSC 122750 and the analogue thereof that produces them, such as 17-AAG.

The invention brief introduction

The invention provides the reduction form of the Ansamycin that comprises benzoquinones, the salt in its unpack format and the pharmaceutical prepn uses their treatments or the adjusting disease relevant with hyper-proliferative such as method for cancer.Usually, the invention provides solvable, the stable medicament forms of the Ansamycin that comprises benzoquinones.The invention provides the reduction analogue of the Ansamycin that contains benzoquinones, such as the amino analogue of the NSC 122750 17-in unpack format and the pharmaceutical prepn, wherein benzoquinones is reduced to quinhydrones and is captured the unpack format into air-stable, such as HCl or H ₂SO ₄Salt.Alternatively, this quinhydrones can be captured and is the common salt of amino acid such as glycocoll.That these analogues have is water-soluble significantly (solvability is than the high 1-3 one magnitude of non-reduced form, and for example 17-AAG is 35 μ g/mL, and the 17-AAG quinhydrones is 1-3mg/mL, and the salt of 17-AAG hydroquinone derivatives is＞200mg/mL) and stability; They can separate and can prepare and be used for human administration and do not have and the non-reduced parent form of Ansamycin and the relevant problem of preparation, storage and unstable of other preparation.

In one embodiment, the invention provides pure isolating formula 1 compound:

Or its free alkali,

Wherein each variable independently is:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

The R of each time appearance independently is selected from hydrogen, alkyl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl;

R ₁Be hydroxyl, alkoxyl group ,-OC (O) R ₈,-OC (O) OR ₉,-OC (O) NR ₁₀R ₁₁,-OSO ₂R ₁₂,-OC (O) NHSO ₂NR ₁₃R ₁₄,-NR ₁₃R ₁₄Or halogen; And R ₂Be hydrogen, alkyl or aralkyl; Or R ₁And R ₂Carbon with their institutes keys connect represents-(C=O)-,-(C=N-OR)-,-(C=N-NHR)-or-(C=N-R)-;

R ₃And R ₄Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆Or R ₃With R ₄Represent the optional substituted heterocycle of 4-8 unit together;

R ₅Be selected from H, alkyl, aralkyl and formula 1a group:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group (acylthio), carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical;

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

R ₈Be hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

R ₉Be alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

R ₁₀And R ₁₁Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆Or R ₁₀And R ₁₁Nitrogen with their institute's keys connect is represented the optional substituted heterocycle of 4-8 unit;

R ₁₂Be alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

R ₁₃And R ₁₄Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆Or R ₁₃And R ₁₄Nitrogen with their institute's keys connect is represented the optional substituted heterocycle of 4-8 unit;

The R of each time appearance ₁₆Independently be selected from hydrogen, hydroxyl, acyl amino,

-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R ₁₈) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉) ,-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) (R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

The R of each time appearance ₁₈Independently be selected from hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl;

The R of each time appearance ₁₉Independently be selected from hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl; Or R ₁₈With R ₁₉Represent the optional substituted ring of 4-8 unit together;

The R of each time appearance ₂₀, R ₂₁, R ₂₂, R ₂₄And R ₂₅It independently is alkyl;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

The R of each time appearance ₂₆And R ₂₇Independently be selected from hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl;

Condition is to work as R ₁Be hydroxyl, R ₂Be hydrogen, R ₆And R ₇Form two keys together, R ₂₀Be methyl, R ₂₁Be methyl, R ₂₂Be methyl, R ₂₃Be methyl, R ₂₄Be methyl, R ₂₅Be methyl, R ₂₆Be hydrogen, R ₂₇Be hydrogen, Q is a key, and W is when being oxygen; R ₃And R ₄Not hydrogen entirely, also do not represent unsubstituted azetidine when combining; And

The absolute stereo chemistry of formula 1 three-dimensional center can be R or S or its mixing, and the stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the invention provides the pure isolated compound that has suc as formula the chemistry of absolute stereo shown in 2:

Or its free alkali;

Wherein each variable independently is:

X ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

R ₁Be hydroxyl or-OC (O) R ₈

R ₃And R ₄Be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆Or R ₃With R ₄Represent the optional substituted heterocycle of 4-8 unit together;

R ₅Be hydrogen or have formula 1a structure:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉), N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical;

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

R ₂₇Be hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl or heteroaralkyl; Condition is to work as R ₁Be hydroxyl, R ₂Be hydrogen, R ₆And R ₇Form two keys together, R ₂₇When being hydrogen; R ₃And R ₄Not hydrogen entirely, also do not represent unsubstituted azetidine when combining; And

The stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the invention provides the pure isolated compound that has suc as formula the chemistry of absolute stereo shown in 3:

X wherein ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root and 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

In another embodiment, the invention provides formula 4 compounds:

Or its pharmacy acceptable salt, wherein each variable independently is:

W is oxygen or sulphur;

Z is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

N equals 0,1 or 2;

M equals 0,1 or 2;

X and Y independently are C (R ₃₀) ₂The R of each time appearance wherein ₃₀Independently be selected from hydrogen, alkyl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl separately; Or-[(CR ₂) _p]-R ₁₆

R ₁Be hydroxyl, alkoxyl group ,-OC (O) R ₈,-OC (O) OR ₉,-OC (O) NR ₁₀R ₁₁,-OSO ₂R ₁₂,-OC (O) NHSO ₂NR ₁₃R ₁₄, NR ₁₃R ₁₄Or halogen; And R ₂Be hydrogen, alkyl or aralkyl; Or R ₁And R ₂Carbon with their institutes keys connect represents-(C=O)-,-(C=N-OR)-,-(C=N-NHR)-or-(C=N-R)-;

R ₃Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆

R ₄Be selected from H, alkyl, aralkyl and formula 4a group:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical;

R ₅And R ₆All be hydrogen; Or R ₅And R ₆Form key together;

-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R ₁₈) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉), ,-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) (R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

The R of each time appearance ₂₆And R ₂₇Independently be selected from hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl; And

The absolute stereo chemistry at formula 4 three-dimensional centers can be R or S or its mixing, and the stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the invention provides the compound that has suc as formula the chemistry of absolute stereo shown in 5:

Wherein each variable independently is:

N equals 0,1 or 2;

M etc. 0,1 or 2

X and Y independently are C (R ₃₀) ₂The R of each time appearance wherein ₃₀Independently be selected from hydrogen, alkyl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl and heteroaralkyl; Or-[(CR ₂) _p]-R ₁₆

R ₁Be hydroxyl or-OC (O) R ₈

R ₃Be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

R ₅Or R ₆All be hydrogen; Or R ₅With R ₆Form key together;

-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R18) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉) ,-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) (R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

R ₂₇Be hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl or heteroaralkyl; And the stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the invention provides and be selected from following compound:

and?

In another embodiment, the present invention provides a kind of pharmaceutical composition, contains: at least a pharmaceutically acceptable vehicle; And formula 6 compounds:

Or its free alkali;

Wherein each variable independently is selected from:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X is the conjugate base of pharmaceutical acceptable acid;

R ₅The group that is selected from H, alkyl, aralkyl and has formula 6a structure:

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

R ₁₀And R ₁₁Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆Or

R ₁₀And R ₁₁Nitrogen with their institute's keys connect is represented the optional substituted heterocycle of 4-8 unit;

R ₁₃Or R ₁₄Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆Or R ₁₃And R ₁₄Nitrogen with their institute's keys connect is represented the optional substituted heterocycle of 4-8 unit;

The R of each time appearance ₁₆Independently be selected from hydrogen, hydroxyl, acyl amino ,-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R ₁₈) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉) ,-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) (R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

The R of each time appearance ₁₈Independently be selected from hydrogen, alkyl, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl or heteroaralkyl;

R for each time appearance ₂₀, R ₂₁, R ₂₂, R ₂₄And R ₂₅It independently is alkyl;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

The absolute stereo chemistry of formula 6 three-dimensional centers can be R or S or its mixing, and the stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the invention provides a kind of pharmaceutical composition, contain: at least a pharmaceutically acceptable vehicle; Formula 6 compounds:

Or its free alkali; With formula 10 compounds, the content of wherein said formula 10 compounds about 0.00001% to about 5% (w/v) scope:

Or its pharmacologically acceptable salt;

Wherein each variable independently is selected from:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

R ₃And R ₄Independently be selected from separately hydrogen, alkyl, alkenyl, alkynyl group, aryl, naphthenic base, Heterocyclylalkyl, aralkyl, heteroaryl, heteroaralkyl and-[(CR ₂) _p]-R ₁₆, or R ₃With R ₄Represent the optional substituted heterocycle of 4-8 unit together;

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

The absolute stereo chemistry of

formula

6 or 10 three-dimensional centers can be R or S or its mixing, and the stereochemistry of two keys can be E or Z or its mixing.

In another embodiment, the present invention relates to a kind of treatment method for cancer, comprise the The compounds of this invention of the Mammals treatment significant quantity that needs this treatment, or the pharmaceutical composition of the present invention of treatment significant quantity.

The present invention relates to the method for preparing compound on the other hand, comprising: in reaction solvent hybrid 7 compounds and reductive agent with production 8 compounds,

and

Mix said formula 8 compounds and pharmaceutically acceptable acid to generate said formula 1 compound;

Wherein each variable independently is:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

R ₅Be selected from H, alkyl, aralkyl and formula 1a group:

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R ₁₈) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉)-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) (R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

Brief description

Accompanying drawing 1 has described to be total to according to the 17-AAG quinhydrones dimethylamino acetate of described method preparation among the embodiment 3 color atlas of the lcms analysis of salt.

Accompanying drawing 2 has described to be total to according to the 17-AAG quinhydrones dimethylamino acetate of described method preparation among the embodiment 3 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones α-An Jiyidingsuan that accompanying drawing 3 has described to prepare according to described method among the embodiment 4 is total to salt ¹H NMR spectrum.

Accompanying drawing 4 has described to be total to according to the 17-AAG quinhydrones α-An Jiyidingsuan of described method preparation among the embodiment 4 color atlas of the lcms analysis of salt.

Accompanying drawing 5 has described to be total to according to the 17-AAG quinhydrones α-An Jiyidingsuan of described method preparation among the embodiment 4 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones Beta-alanine that accompanying drawing 6 has described to prepare according to described method among the embodiment 5 is total to salt ¹H NMR spectrum.

Accompanying drawing 7 has described to be total to according to the 17-AAG quinhydrones Beta-alanine of described method preparation among the embodiment 5 color atlas of the lcms analysis of salt.

Accompanying drawing 8 has described to be total to according to the 17-AAG quinhydrones Beta-alanine of described method preparation among the embodiment 5 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones sarcosine that accompanying drawing 9 has described to prepare according to described method among the embodiment 6 is total to salt ¹H NMR spectrum.

Accompanying drawing 10 has described to be total to according to the 17-AAG quinhydrones sarcosine of described method preparation among the embodiment 6 color atlas of the lcms analysis of salt.

Accompanying drawing 11 has described to be total to according to the 17-AAG quinhydrones sarcosine of described method preparation among the embodiment 6 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones piperidine carboxylic acid that accompanying drawing 12 has described to prepare according to described method among the embodiment 7 is total to salt ¹H NMR spectrum.

Accompanying drawing 13 has described to be total to according to the 17-AAG quinhydrones piperidine carboxylic acid of described method preparation among the embodiment 7 color atlas of the lcms analysis of salt.

Accompanying drawing 14 has described to be total to according to the 17-AAG quinhydrones piperidine carboxylic acid of described method preparation among the embodiment 7 mass spectrum of the lcms analysis of salt.

Accompanying drawing 15 has described to be total to according to the 17-AAG quinhydrones piperidine carboxylic acid of described method preparation among the embodiment 7 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones glycocoll that accompanying drawing 16 has described to prepare according to described method among the embodiment 8 is total to salt ¹H NMR spectrum.

Accompanying drawing 17 has described to be total to according to the 17-AAG quinhydrones glycocoll of described method preparation among the embodiment 8 color atlas of the lcms analysis of salt.

Accompanying drawing 18 has described to be total to according to the 17-AAG quinhydrones glycocoll of described method preparation among the embodiment 8 mass spectrum of the lcms analysis of salt.

17-AAG quinhydrones 2-amino-2-ethyl-butyric acid that accompanying drawing 19 has described to prepare according to described method among the embodiment 9 is total to salt ¹H NMR spectrum.

Accompanying drawing 20 has described to be total to according to the 17-AAG quinhydrones 2-amino-2-ethyl-butyric acid of described method preparation among the embodiment 9 color atlas of the lcms analysis of salt.

Accompanying drawing 21 has described to be total to according to the 17-AAG quinhydrones 2-amino-2-ethyl-butyric acid of described method preparation among the embodiment 9 mass spectrum of the lcms analysis of salt.

17-AAG quinhydrones 1-amino-cyclopropane-carboxylic acid that accompanying drawing 22 has described to prepare according to described method among the embodiment 10 is total to salt ¹H NMR spectrum.

Accompanying drawing 23 has described to be total to according to the 17-AAG quinhydrones 1-amino-cyclopropane-carboxylic acid of described method preparation among the embodiment 10 color atlas of the lcms analysis of salt.

Accompanying drawing 24 has described to be total to according to the 17-AAG quinhydrones 1-amino-cyclopropane-carboxylic acid of described method preparation among the embodiment 10 mass spectrum of the lcms analysis of salt.

The 17-AAG hydroquinone carboxylic acid that accompanying drawing 25 has described to prepare according to described method among the embodiment 11 is total to salt ¹H NMR spectrum.

Accompanying drawing 26 has described to be total to according to the 17-AAG hydroquinone carboxylic acid of described method preparation among the embodiment 11 color atlas of the lcms analysis of salt.

Accompanying drawing 27 has described to be total to according to the 17-AAG hydroquinone carboxylic acid of described method preparation among the embodiment 11 mass spectrum of the lcms analysis of salt.

17-AAG quinhydrones 1-amino-Cyclopentane carboxylic acid that accompanying drawing 28 has described to prepare according to described method among the embodiment 12 is total to salt ¹H NMR spectrum.

Accompanying drawing 29 has described to be total to according to the 17-AAG quinhydrones 1-amino-Cyclopentane carboxylic acid of described method preparation among the embodiment 12 color atlas of the lcms analysis of salt.

Accompanying drawing 30 has described to be total to according to the 17-AAG quinhydrones 1-amino-Cyclopentane carboxylic acid of described method preparation among the embodiment 12 mass spectrum of the lcms analysis of salt.

The 17-AAG quinhydrones N-methyl piperidine carboxylic acid that accompanying drawing 31 has described to prepare according to described method among the embodiment 13 is total to salt ¹H NMR spectrum.

Accompanying drawing 32 has described to be total to according to the 17-AAG quinhydrones N-methyl piperidine carboxylic acid of described method preparation among the embodiment 13 color atlas of the lcms analysis of salt.

Accompanying drawing 33 has described to be total to according to the 17-AAG quinhydrones N-methyl piperidine carboxylic acid of described method preparation among the embodiment 13 mass spectrum of the lcms analysis of salt.

Accompanying drawing 34 has been described the 17-AAG quinhydrones N according to described method preparation among the embodiment 14, N, the common salt of N-trimethyl ammonium acetate ¹H NMR spectrum.

Accompanying drawing 35 has been described according to the 17-AAG hydroquinone derivatives of the air of described method preparation among the embodiment 15-stable ¹H NMR spectrum.

Accompanying drawing 36 has been described according to the HCl salt of the 17-AAG hydroquinone derivatives of described method preparation among the embodiment 17 ¹H NMR spectrum.

Accompanying drawing 37 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones HCl salt of described method preparation among the embodiment 17.

Accompanying drawing 38 has been described the 17-AAG hydroquinone derivatives H according to described method preparation among the embodiment 18 ₂SO ₄Salt ¹H NMR spectrum.

Accompanying drawing 39 has been described the 17-AAG quinhydrones H according to described method preparation among the embodiment 18 ₂SO ₄The color atlas of the lcms analysis of salt.

Accompanying drawing 40 has been described the 17-AAG quinhydrones H according to described method preparation among the embodiment 18 ₂SO ₄The mass spectrum of the lcms analysis of salt.

Accompanying drawing 41 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones tosilate of described method preparation among the embodiment 19.

Accompanying drawing 42 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones tosilate of described method preparation among the embodiment 19.

Accompanying drawing 43 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones tosilate of described method preparation among the embodiment 19.

Accompanying drawing 44 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones d-camsilate of described method preparation among the embodiment 20.

Accompanying drawing 45 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones d-camsilate of described method preparation among the embodiment 20.

Accompanying drawing 46 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones d-camsilate of described method preparation among the embodiment 20.

Accompanying drawing 47 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones d-camsilate of described method preparation among the embodiment 20.

Accompanying drawing 48 has been described the 17-AAG quinhydrones H according to described method preparation among the embodiment 21 ₃PO ₄The color atlas of the lcms analysis of salt.

Accompanying drawing 49 has been described the 17-AAG quinhydrones H according to described method preparation among the embodiment 21 ₃PO ₄The mass spectrum of the lcms analysis of salt.

Accompanying drawing 50 has been described the 17-AAG quinhydrones MeSO according to described method preparation among the embodiment 22 ₃The color atlas of the lcms analysis of H salt.

Accompanying drawing 51 has been described the 17-AAG quinhydrones MeSO according to described method preparation among the embodiment 22 ₃The mass spectrum of the lcms analysis of H salt.

Accompanying drawing 52 has been described the 17-AAG quinhydrones PhSO according to described method preparation among the embodiment 23 ₃The color atlas of the lcms analysis of H salt.

Accompanying drawing 53 has been described the 17-AAG quinhydrones PhSO according to described method preparation among the embodiment 23 ₃The mass spectrum of the lcms analysis of H salt.

Accompanying drawing 54 has been described according to the 17-AAG quinhydrones cyclic amino formate of described method preparation among the embodiment 25 ¹H NMR spectrum.

Accompanying drawing 55 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones cyclic amino formate of described method preparation among the embodiment 25.

Accompanying drawing 56 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones cyclic amino formate of described method preparation among the embodiment 25.

Accompanying drawing 57 has been described according to the 17-AAG quinhydrones lactan of described method preparation among the embodiment 26 ¹H NMR spectrum.

Accompanying drawing 58 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones lactan of described method preparation among the embodiment 26.

Accompanying drawing 59 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones lactan of described method preparation among the embodiment 26.

Accompanying drawing 60 has been described according to the 17-aminoderivative of the 17-AAG quinhydrones of described method preparation among the embodiment 27 ¹H NMR spectrum.

Accompanying drawing 61 has been described the color atlas according to the lcms analysis of the 17-aminoderivative of the 17-AAG quinhydrones of described method preparation among the embodiment 27.

Accompanying drawing 62 has been described the mass spectrum according to the lcms analysis of the 17-aminoderivative of the 17-AAG quinhydrones of described method preparation among the embodiment 27.

Accompanying drawing 63 has been described according to 17-(3-amino-propane-1, the 2-glycol) verivate of the 17-AAG quinhydrones of described method preparation among the embodiment 28 ¹H NMR spectrum.

Accompanying drawing 64 has been described the color atlas according to the lcms analysis of 17-(3-amino-propane-1, the 2-glycol) verivate of the 17-AAG quinhydrones of described method preparation among the embodiment 28.

Accompanying drawing 65 has been described the mass spectrum according to the lcms analysis of 17-(3-amino-propane-1, the 2-glycol) verivate of the 17-AAG quinhydrones of described method preparation among the embodiment 28.

Accompanying drawing 66 has been described the color atlas according to the lcms analysis of the BODIPY verivate of the 17-AAG quinhydrones of described method preparation among the embodiment 29.

Accompanying drawing 67 has been described the mass spectrum according to the lcms analysis of the BODIPY verivate of the 17-AAG quinhydrones of described method preparation among the embodiment 29.

Accompanying drawing 68 has been described according to the 17-AAG quinhydrones HBr salt of described method preparation among the embodiment 31 ¹H NMR spectrum.

Accompanying drawing 69 has been described the color atlas according to the lcms analysis of the 17-AAG quinhydrones HBr salt of described method preparation among the embodiment 31.

Accompanying drawing 70 has been described the mass spectrum according to the lcms analysis of the 17-AAG quinhydrones HBr salt of described method preparation among the embodiment 31.

Detailed Description Of The Invention

General introduction

The present invention relates to pure isolating reduction form analogue, its salt and the midbody that comprises the Ansamycin of benzoquinones.The present invention also provides disease or the state of an illness such as cancer and the disease relevant with unwanted HSP90 activity or wherein HSP90 onset in relating to the cell that causes disorder or the method for the state of an illness of using these compounds for treating to be characterised in that the cell proliferation of not expecting.The invention provides the reduction analogue that comprises the benzoquinones Ansamycin, wherein benzoquinones is reduced to quinhydrones, and preferred separated purifying is a salt form.In a kind of alternative embodiment, The compounds of this invention and amino acid salts cocrystallization.These analogues have or do not have amino acid salts, have all that water-soluble significantly (than the high 1-3 one magnitude of non-reduced form, for example 17-AAG is 35 μ g/mL, and the 17-AAG quinhydrones is 1-3mg/mL, and quinhydrones salt is＞200mg/mL) and at room temperature stable; They can separate and can prepare and be used for human administration and do not have and the non-reduced parent form of Ansamycin and the relevant problem of preparation, storage and unstable of other preparation.

Definition

The definition of term used herein is intended to comprise that present this area that chemistry and pharmaceutical field are generally acknowledged for each term defines.In the time of suitably, instance is provided.Only if restriction is arranged in object lesson in addition, these definition are individually or as than the certain applications of macoradical in this specification sheets in full.

When specifically not indicating stereochemistry, all steric isomers of The compounds of this invention comprise all that within the scope of the present invention it is pure compound or its mixture.Unless otherwise, each enantiomorph, diastereomer, geometrical isomer with and combination and mixture include in the present invention.Polymorphic and solvolyte are also included within the scope of the invention.

As used herein, term " amino acid " is meant and not only comprises carboxylic moiety but also comprise amino molecule partly.Carboxylic acid and amino part are as giving a definition.Naturally occurring and synthetic deutero-amino acid includes within the scope of the present invention.

As described herein; Term " benzoquinones Ansamycin " is meant the compound that contains Macrocyclic lactams; In ring, only contain a lactan and in lactam nucleus, contain the benzoquinones part further; Wherein said benzoquinones partly has at least one nitrogen substituting group, and one in wherein said at least one nitrogen substituting group is the part of an only amide moieties in the said lactan.The object lesson that can be used for naturally occurring benzoquinones Ansamycin of the present invention includes but not limited to, NSC 122750 and herbimycin.Term " NSC 122750 analogue " is meant and can passes through for example chemically modified and deutero-benzoquinones Ansamycin by NSC 122750; For example 17-allyl amino-17-demethoxylation NSC 122750 (17-AAG) or 17-(2-dimethyl aminoethyl) amino-17-demethoxylation NSC 122750 (17-DMAG).

As used herein, the term that links to each other with The compounds of this invention " isolating " means compound not in cell or organism, but compound is separated with common natural its some or all components of following.

As used herein, the term " pure " that links to each other with the isolating sample of The compounds of this invention means the compound that isolating sample comprises at least 60% weight.Preferably, isolating sample comprises the compound of at least 70% weight.More preferably, isolating sample comprises the compound of at least 80% weight.More preferably, isolating sample comprises the compound of at least 90% weight.Most preferably, isolating sample comprises the compound of at least 95% weight.The purity of the isolating sample of The compounds of this invention can be estimated through many methods or its combination, for example: thin layer, preparation or flash chromatography, mass spectrum, HPLC, NMR analysis etc.

Term " heteroatoms " is the cognitive any element of also finger except carbon or hydrogen in this area.The heteroatoms of exemplary comprises boron, nitrogen, oxygen, phosphorus, sulphur and selenium.

Term " alkyl " be this area cognitive and comprise saturated aliphatic group, comprise the alkyl of straight chained alkyl, branched-chain alkyl, cycloalkanes (alicyclic ring) base, the substituted naphthenic base of alkyl and cycloalkyl substituted.In certain embodiments, the straight or branched alkyl has about 30 or carbon atom still less (for example straight chain is C1-C30, and side chain is C3-C30) in its skeleton, and alternatively, and about 20 or still less.Likewise, naphthenic base has about 10 carbon atoms of about 3-in its ring structure, and about 5,6 or 7 carbon in ring structure alternatively.

Only if the appointment carbon number is arranged in addition, " low alkyl group " is meant the as above alkyl of definition, but in its skeleton structure, has about 10 carbon of 1-, alternatively about 6 carbon of 1-.Likewise " low-grade alkenyl " has similar chain length with " alkynyl of low-grade chain ".

Term " aralkyl " is cognitive also the referring to by the substituted alkyl of aryl (for example, aryl or heteroaryl) in this area.

Term " alkenyl " and " alkynyl group " be this area cognitive and refer to that length and possible substituting group are similar to the unsaturated aliphatic group of abovementioned alkyl, but comprise at least one two keys or triple bond respectively.

Term " aryl " be this area cognitive and refer to 5-, 6-and 7-unit monocyclic aryl; It can comprise 0-4 heteroatoms; For example, benzene, naphthalene, anthracene, pyrene, pyrroles, furans, thiophene, imidazoles, _ azoles, thiazole, triazole, pyrazoles, pyridine, pyrazine, pyridazine and pyrimidine, or the like.In ring structure, have heteroatomic those aryl and also can be called as " fragrant heterocycle " or " assorted aromatic hydrocarbons ".Aromatic ring can be replaced by above-mentioned substituting group at one or more ring positions, for example halogen atom, nitrine, alkyl, aralkyl, alkenyl, alkynyl group, naphthenic base, hydroxyl, alkoxyl group, amino, nitro, sulfydryl, imino-, amido, phosphonate group, phosphonous acid base, carbonyl, carboxyl, silyl, ether, alkylthio, alkylsulfonyl, sulfoamido, ketone, aldehyde, ester, heterocyclic radical, virtue or assorted virtue part ,-CF ₃,-CN, or the like.Term " aryl " also comprises the many rings ring system with two or more rings; Wherein two or more carbon are two, and the chain of rings (this ring is " fused rings ") is total mutually; Wherein at least one ring is fragrant, and for example other ring can be naphthenic base, cycloalkenyl group, cycloalkynyl radical, aryl and/or heterocyclic radical.

Term Adjacent, BetweenWith RightBe this area cognitive and refer to 1 respectively, 2-, 1,3-and 1,4-disubstituted benzenes.For example, 1, the 2-dimethyl benzene with Adjacent-YLENE synonym.

Term " heterocyclic radical ", " heteroaryl " or " heterocyclic group " be this area cognitive and refer to 3-to about 10-ring structure, alternatively 3-encircles to about 7-is first, its ring structure comprises 1-4 heteroatoms.Heterocycle can also be many rings.Heterocycle for example comprises; Thiophene, thianthrene, furans, pyrans, isobenzofuran, chromene, xanthene, fen cluck ton, pyrroles, imidazoles, pyrazoles, isothiazole, different _ azoles, pyridine, pyrazine, pyrimidine, pyridazine, pyrrocoline, isoindole, indoles, indazole, purine, quinolizine, isoquinoline 99.9, quinoline, phthalazines, 1.5-naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridines, acridine, pyrimidine, phenanthroline, azophenlyene, phenarsazine, thiodiphenylamine, furazan, fen _ piperazine, tetramethyleneimine, oxolane, thiolane, _ azoles, piperidines, piperazine, morpholine, lactone, lactan be such as azetidinone and pyrrolidone, sultam, sultone, or the like.Heterocycle can be replaced by above-mentioned substituting group in one or more positions, for example halogen atom, nitrine, alkyl, aralkyl, alkenyl, alkynyl group, naphthenic base, hydroxyl, alkoxyl group, amino, nitro, sulfydryl, imino-, amido, phosphonate group, phosphonous acid base, carbonyl, carboxyl, silyl, ether, alkylthio, alkylsulfonyl, ketone, aldehyde, ester, heterocyclic radical, virtue or assorted virtue part ,-CF ₃,-CN, or the like.

Term " optional replace " is meant chemical group such as alkyl, naphthenic base aryl etc., wherein one or more hydrogen by substituting group as described herein for example halogen atom, nitrine, alkyl, aralkyl, alkenyl, alkynyl group, naphthenic base, hydroxyl, alkoxyl group, amino, nitro, sulfydryl, imino-, amido, phosphonate group, phosphonous acid base, carbonyl, carboxyl, silyl, ether, alkylthio, alkylsulfonyl, sulfoamido, ketone, aldehyde, ester, heterocyclic radical, virtue or assorted virtue partly ,-CF ₃,-CN etc. substitute.

Term " many ring " or " many cyclic groups " be this area cognitive and refer to two or more rings (for example, naphthenic base, cycloalkenyl group, cycloalkynyl radical, aryl and/or heterocyclic radical), wherein two or more carbon be two a chain of mutually total, as, this ring is " fused rings ".The ring that connects through non-adjacent atom is called as " bridged ring ".Each ring of polycyclic can be replaced by above-mentioned substituting group, for example halogen atom, alkyl, aralkyl, alkenyl, alkynyl group, naphthenic base, hydroxyl, amino, nitro, sulfydryl, imino-, amido, phosphonate group, phosphonous acid base, carbonyl, carboxyl, silyl, ether, alkylthio, alkylsulfonyl, ketone, aldehyde, ester, heterocyclic radical, virtue or assorted virtue partly ,-CF ₃,-CN, or the like.

Term " carbocyclic ring " is that this area is cognitive, and refers to that wherein each annular atoms is the fragrance or the non-aromatic ring of carbon.

Term " nitro " is the cognitive also finger-NO in this area ₂Term " halogen atom " be the cognitive also finger-F in this area ,-Cl ,-Br or-I; Term " sulfydryl " is the cognitive also finger-SH in this area; Term " hydroxyl " means-OH; And term " alkylsulfonyl " is the cognitive also finger-SO in this area ₂ ^-The respective anionic of " halogenide " expression halogen atom, and " false halogenide (pseudohalide) " has 560 the above definition of " Advanced InorganicChemistry " by Cotton andWilkinson.

Term " amine " and " amino " be this area cognitive also refer to not replacement and substituted amine, the part that for example can represent by following general formula:

R wherein ₅₀, R ₅₁And R ₅₂Independent separately represent hydrogen, alkyl, alkenyl ,-(CH ₂) _m-R ₆₁Or R ₅₀And R ₅₁, constitute ring structure with 4-8 atom with the N atom that they connected; R ₆₁Represent aryl, naphthenic base, cycloalkenyl group, heterocycle or many rings; M is 0 or the integer of 1-8.In other embodiments, R ₅₀And R ₅₁(and optional R ₅₂) independent separately represent hydrogen, alkyl, alkenyl or-(CH ₂) _m-R ₆₁Thus, term " alkylamine " comprises as above the amine groups of definition, has the replacement that is attached thereto or substituted alkyl not, that is, and and at least one R ₅₀And R ₅₁It is alkyl.

Term " acyl amino " is the part that the cognitive also finger in this area can be represented by formula:

R wherein ₅₀As above definition, and R ₅₄Represent hydrogen, alkyl, alkenyl or-(CH ₂) _m-R ₆₁, wherein m and R ₆₁As above definition.

Term " carboxamido-group " is the cognitive amino substituted carbonyl in this area, and comprises the part that can be represented by formula:

R wherein ₅₀And R ₅₁As above definition.Some embodiment of acid amides of the present invention will not comprise the imide of potentially unstable.

Term " alkylthio (alkylthio) " is meant as above the alkyl of definition, and it is coupled to have a methylthio group.In certain embodiments, " alkylthio " part by-S-alkyl ,-the S-alkenyl ,-the S-alkynyl group and-S-(CH ₂) _m-R ₆₁One of representative, wherein m and R ₆₁As above definition.Representative alkylthio comprises methylthio group, ethylmercapto group etc.

Term " carboxyl " be this area cognitive and comprise can be by the part of formula representative:

X wherein ₅₀Be key or represent oxygen or sulphur, and R ₅₅And R ₅₆Represent hydrogen, alkyl, alkenyl ,-(CH ₂) _m-R ₆₁Or pharmacy acceptable salt, R ₅₆Represent hydrogen, alkyl, alkenyl or-(CH ₂) _m,-R ₆₁, wherein m and R ₆₁As above definition.X wherein ₅₀Be oxygen and R ₅₅Or R ₅₆When being not hydrogen, this formula representative " ester ".X wherein ₅₀Be oxygen, and R ₅₅When as above defining, this part refers to carboxyl herein, and especially, works as R ₅₅When being hydrogen, this formula representative " carboxylic acid ".X wherein ₅₀Be oxygen, and R ₅₆Be hydrogen, this formula representative " manthanoate " is common, when the following formula Sauerstoffatom is substituted by sulphur, and this formula representative " thiocarbonyl ".X wherein ₅₀Be sulphur and R ₅₅Or R ₅₆When being not hydrogen, this formula representative " monothioester ".X wherein ₅₀Be sulphur and R ₅₅When being hydrogen, this formula representative " thiocarboxylic acid ".X wherein ₅₀Be sulphur and R ₅₆When being hydrogen, this formula representative " thiocarboxylic ".On the other hand, X wherein ₅₀Be key, and R ₅₅Not hydrogen, following formula representative " ketone group ".X wherein ₅₀Be key, and R ₅₅Be hydrogen, following formula representative " aldehyde " base.

Term " formamyl " is meant-O (C=O) NRR ' that wherein R and R ' independently are H, aliphatic group, aryl or heteroaryl.

Term " oxo " be meant ketonic oxygen (=O).

Term " oxime " and " oxime ether (oxime ether) " are that this area is cognitive, and comprise can be by the part of formula representative:

R wherein ₇₅Be hydrogen, alkyl, naphthenic base, alkenyl, alkynyl group, aryl, aralkyl or-(CH ₂) _m-R ₆₁This part is " oxime " when R is H; When R be alkyl, naphthenic base, alkenyl, alkynyl group, aryl, aralkyl or-(CH ₂) _m-R ₆₁The time it is " an oxime ether ".

Term " alkoxyl group " or " alcoxyl " be this area approval also refer to the alkyl that as above defines, it is coupled to have an oxygen groups.Representational alkoxyl group comprises methoxyl group, oxyethyl group, propoxy-, tert.-butoxy, or the like." ether " is that two hydrocarbon and oxygen are covalently bound.Therefore, make that alkyl is that the alkyl substituent of ether is or is similar to alkoxyl group, as can by-O-alkyl ,-the O-alkenyl ,-the O-alkynyl group ,-O--(CH ₂) _m-R ₆₁One of representative, wherein m and R ₆₁As above definition.

Term " sulfonic group " be this area approval also refer to the part that can represent by formula:

R wherein ₅₇Be electron pair, hydrogen, alkyl, naphthenic base or aryl.

Term " sulfate " be this area approval also refer to the part that can represent by formula:

R wherein ₅₇As above definition.

Term " sulfoamido " be this area approval also refer to the part that can represent by formula:

R wherein ₅₀And R ₅₆As above definition.

Term " sulfamyl " be this area approval also refer to the part that can represent by formula:

R wherein ₅₀And R ₅₁As above definition.

Term " alkylsulfonyl " be this area approval also refer to the part that can represent by formula:

R wherein ₅₈Be one of following: hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, heterocyclic radical, aryl or heteroaryl.

Term " sulfoxide group (sulfoxido) " be this area approval also refer to the part that can represent by formula:

R wherein ₅₈As above definition.

Term " phosphoryl " be this area approval also refer to the part that can represent by formula:

Wherein Q50 represents S or O, and R ₅₉Represent hydrogen, low alkyl group or aryl.

When for example being used to replace alkyl, the phosphoryl of phosphoryl alkyl can be represented by formula:

Wherein Q50 and R ₅₉Independent separately as above definition, and Q51 represents O, S or N.When Q50 was S, this phosphoryl partly was " thiophosphatephosphorothioate (phosphorothioate) ".

Term " phosphoramidate (phosphoramidite) " is also can being represented by formula of this area approval:

Wherein Q51, R ₅₀, R ₅₁And R ₅₉As above definition.

Term " amido phosphonate (phosphonamidite) " is also can being represented by formula of this area approval:

Wherein Q51, R ₅₀, R ₅₁And R ₅₉As above definition, and R ₆₀Represent low alkyl group or aryl.

Can similarly replace alkenyl and alkynyl group, to form for example amino alkenyl, amino alkynyl group, amido alkenyl, amido alkynyl group, imino-alkenyl, imino-alkynyl group, sulfo-alkenyl, sulfo-alkynyl group, carbonyl-substituted alkenyl or alkynyl group.

When each express as alkyl, m, n etc. be defined in that appearance is more than one time in any structure time, its mean with same structure in the definition of other position separate.

Term " seleno alkyl " is that the also finger of this area approval has the coupled alkyl of substituted seleno.Can be on alkyl substituted representational " seleno ether " be selected from-the Se-alkyl ,-the Se-alkenyl ,-the Se-alkynyl group and-Se-(CH ₂) _m-R ₆₁, m and R ₆₁As above definition.

Term triflyl, tosyl, mesyl and nonaflyl are that this area is approved and refer to trifyl, p-toluenesulfonyl, methylsulfonyl and nine fluorine fourth alkylsulfonyls respectively.Term triflate, tosylate, mesylate and nonaflate be this area approval and refer to fluoroform sulphonyl acid esters, tolysulfonyl acid esters, methanesulfonates and nine fluorine fourth sulfonate functionality respectively and comprise the molecule of said group.

Abbreviation Me, Et, Ph, Tf, Nf, Ts and Ms be represent methylidene, ethyl, phenyl, trifyl, nine fluorine fourth alkylsulfonyls, p-toluenesulfonyl and methylsulfonyl respectively.The more comprehensively tabulation of the used abbreviation of organic chemistry filed technician is shown in the chapter 1 of each volume of Journal of OrganicChemistry; This tabulation is generally by entitling The abbreviation standard listThe form of (Standard List of Abbreviations).

Some compound that is contained in the present composition can exist with concrete geometry or stereoisomer form.In addition, polymkeric substance of the present invention can also be optically active.It is considered herein that all these compounds, comprise cis-with trans-isomer, R-and S-enantiomorph, diastereomer (D)-isomer, (L)-isomer, its racemic mixture with and other mixture all fall into the scope of the invention.Other unsymmetrical carbon also can be present in substituting group such as the alkyl.All these isomer and composition thereof all are intended within the present invention.

For example, a certain concrete enantiomorph of The compounds of this invention if desired, can derive through asymmetric synthesis or through the chirality assistant agent prepares, and wherein separating obtained mixture of diastereomers also ruptures the assistant agent group and obtains purified required enantiomorph.Alternatively; When molecule comprises basic functionality such as amino, or acidic functionality can form non-mapping salt with appropriate optical live acid or alkali during such as carboxyl; With after fractional crystallization well known in the art or chromatographic process split the diastereomer that forms, and reclaim pure enantiomorph subsequently.

Be interpreted as " replacement " or " being substituted " comprised implied condition; Be that these replacements should be consistent with the valency that substituted atom and substituting group allowed; This replacement causes forming stable compound, and for example it can spontaneous transition of carrying out such as rearrangement, cyclisation, elimination or other reaction.

The substituting group of the organic cpds that term " substituted " is intended to comprise that all are allowed.One widely aspect, the substituting group of allowing includes acyclic and ring-type, side chain and non-side chain, carbocyclic ring and heterocycle, fragrance and the non-aromatic substituent of organic compounds.Representational substituting group for example comprises above-mentioned those.The substituting group of allowing for suitable organic cpds have one or more, identical or different.For the purposes of the present invention, heteroatoms such as nitrogen can have the hydrogen substituting group and/or as herein described any that allow, satisfy the valent organic cpds substituting group of heteroatoms.The present invention is not that the substituting group that is intended to allowed by organic cpds by any way limits.

As used herein, term " protection base " means interim substituting group, and it prevents that the potential reactive functionality from carrying out unwanted chemical conversion.These protection bases for example comprise the ester of carboxylic acid, the silicomethane ether of alcohol and the acetal and the ketal of aldehyde and ketone respectively.The basic chemical field of protection (Greene, T.W have been summarized; Wuts, P. G.M.Protective Groups inOrganicSynthesis, 2 ^NdEd.; Wiley:New York, 1991).The protection form of The compounds of this invention comprises within the scope of the present invention.

For the purposes of the present invention, according to the periodic table of elements, CAS version, Handbook ofChemistry and Physics, 67 ^ThEd., front cover is identified chemical element in the 1986-87.

Term " pharmacy acceptable salt " or " salt " are meant the salt of one or more compounds.The suitable pharmacy acceptable salt of compound comprises acid salt; It for example can form through the solution of mixing cpd solution and pharmaceutical acceptable acid; Pharmaceutical acceptable acid is such as spirit of salt, Hydrogen bromide, sulfuric acid, fumaric acid, toxilic acid, succsinic acid, phenylformic acid, acetate, Hydrocerol A, tartrate, phosphoric acid, carbonic acid, or the like.When compound is loaded with one or more acid moieties; Pharmacy acceptable salt can form through the solution-treated compound solution that uses pharmaceutically acceptable alkali; Pharmaceutically acceptable alkali is such as Lithium Hydroxide MonoHydrate, sodium hydroxide, Pottasium Hydroxide, tetraalkylammonium hydroxide, Quilonum Retard, yellow soda ash, salt of wormwood, ammonia, alkylamine, or the like.

Term " pharmaceutical acceptable acid " is meant the inorganic or organic acid that demonstration is nontoxic basically.Pharmaceutical acceptable acid includes but not limited to spirit of salt, Hydrogen bromide, sulfuric acid, nitric acid, Phenylsulfonic acid, methylsulfonic acid, fumaric acid, toxilic acid, succsinic acid, phenylformic acid, acetate, Hydrocerol A, tartrate, phosphoric acid, carbonic acid, or the like.

Term " salt altogether " or " cocrystallization " are meant the compsn that reductive Ansamycin salt form exists such as amino acid salts with at least a other salt.

As used herein, term " experimenter " is meant animal, normally Mammals or people, its will be or be treatment, observe and/or the object of test.When this term is used for when giving compound or medicine and link to each other, the experimenter is exactly treatment, observe and/or give the object of compound or medicine.

Term " administration altogether " and " giving altogether " are meant administration (giving two or more medicines simultaneously) simultaneously and different time administration (giving one or more other medicines in the time with the asynchronism(-nization) that gives one or more medicines), as long as medicine is present in patient's body to a certain extent simultaneously.

As used herein; Term " treatment significant quantity " means active compound or medicine shows biology or the active compound of drug responses or the amount of medicine that investigator, animal doctor, clinician or doctor investigate in cell culture, tissue system, animal or human, and said replying comprises the symptom of alleviating the disease of treating, the state of an illness or disorder.

Term " compsn " is intended to comprise the product that contains the specified quantitative special component; And any product that directly or indirectly forms by combination specified quantitative special component; Particularly be total to salt, such as reductive Ansamycin salt (like vitriol) and amino acid (like glycocoll) salt.

Term " disorder of HSP90 mediation " or " by the disorder of cell expressing HSP90 mediation " are meant wherein acting pathology of HSP90 and disease condition.This effect can be directly related with pathologic condition or can with state of an illness indirect correlation.The common trait of this type of state of an illness is that this state of an illness can be alleviated with other proteic combination through the activity, function or the HSP90 that suppress HSP90.

Term " pharmaceutically acceptable carrier " is meant the vehicle that is used to prepare the required formulation of compound.Pharmaceutically acceptable carrier can comprise one or more solvents, thinner or other liquid vehicle; Disperse or the suspension assistant agent; Tensio-active agent; Isotonic agent; Thickening material or emulsifying agent; Sanitas; Solid binder; Lubricant; Or the like.Remington ' s PharmaceuticalSciences, Fifteenth Edition, E.W.Martin (Mack Publishing Co., Easton, Pa., 1975) and Handbook of Pharmaceutical Excipients, ThirdEdition, A.H.Kibbe ' ed. (American Pharmaceutical Assoc.2000) openly are used for the variety carrier and the known technology for preparing it of compounding pharmaceutical compsn.

The explanation of some preferred embodiment

The present invention has explained the Ansamycin that needs to produce soluble form, particularly comprises the member of benzoquinones family, like NSC 122750.These types member of macrocycle molecule trends towards very indissoluble, causes being difficult to become potential medicine (for example, the solubleness of 17-AAG in the aqueous solution is merely 100 μ g/mL).The present invention solves these problems through general reaction process is provided, and its reaction process can be used for generating and has the analog molecules of improving solubleness.These reaction process comprise and reduce the quinone of these molecules forming quinhydrones, and capture into salt such as HCl or H ₂SO ₄Salt.Significantly, for example, the quinhydrones HCl salt of 17-AAG has＞solubleness of about 200mg/mL.

Compound

The present invention also provides the separated similar thing of Ansamycin that comprises benzoquinones, and wherein benzoquinones is reduced to quinhydrones and is captured with the reaction of suitably organic or inorganic acid through quinhydrones and is ammonium salt.

In one embodiment, the invention provides pure isolating formula 1 compound:

Or its free alkali, wherein each variable independently is:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

R ₅Be selected from H, alkyl, aralkyl and formula 1a group:

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, condition is to work as R ₁Be hydroxyl, R ₂Be hydrogen, R ₃Be hydrogen, R ₆And R ₇Form two keys together, R ₂₀Be methyl, R ₂₁Be methyl, R ₂₂Be methyl, R ₂₃Be methyl, R ₂₄Be methyl, R ₂₅Be methyl, R ₂₆Be hydrogen, R ₂₇Be hydrogen, Q is a key, and W is when being oxygen; R ₃And R ₄Not hydrogen entirely, also do not represent unsubstituted azetidine when combining.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₂₀, R ₂₁, R ₂₂, R ₂₃, R ₂₄And R ₂₅Be methyl, R ₂₆Be hydrogen, Q is a key; And W is an oxygen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, the pKa of wherein said pharmaceutically acceptable acid in water is about between-10 to about 4.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, the pKa of wherein said pharmaceutically acceptable acid in water is about between-10 to about 1.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, the pKa of wherein said pharmaceutically acceptable acid in water is about-10 to approximately between-3.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein X ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₂Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₃And R ₄Independently be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₅Be hydrogen or have formula 1a structure:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₆And R ₇Form two keys together.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₂₇Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen; R ₃And R ₄Independently be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen; R ₃And R ₄Independently be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆And R ₅Be hydrogen or have formula 1a structure:

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen; R ₃And R ₄Independently be hydrogen, alkyl, alkenyl, alkynyl group, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆R ₅Be hydrogen or have formula 1a structure:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂O-heterocyclic radical, and R ₆And R ₇Form two keys together.

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂O-heterocyclic radical, and R ₆And R ₇Form two keys together; And R ₂₇Be hydrogen.

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂O-heterocyclic radical, and R ₆And R ₇Form two keys together; R ₂₇Be hydrogen; And X ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂O-heterocyclic radical, and R ₆And R ₇Form two keys together; R ₂₇Be hydrogen; And X ^-Be selected from chlorine and bromine.

Or its free alkali;

Wherein each variable independently is:

R ₁Be hydroxyl or-OC (O) R ₈

R ₅Be hydrogen or have formula 1a structure:

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

The stereochemistry of two keys can be E or Z or its mixing.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, condition is to work as R ₁Be hydroxyl, R ₂Be hydrogen, R ₆And R ₇Form two keys together, R ₂₇When being hydrogen; R ₃And R ₄Not hydrogen entirely, also do not represent unsubstituted azetidine when combining.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₃It is allyl group.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₃Have formula 9 structures:

Or its free alkali;

X wherein ₁ ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₄Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₅Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₆And R ₇Form key together.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; And R ₄Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃Have formula 9 structures:

Or its free alkali;

X wherein ₁ ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate. And R ₄Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₄Be hydrogen; And R ₅Be hydrogen.

Or its free alkali; X wherein ₁ ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate. R ₄Be hydrogen; And R ₅Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₄Be hydrogen; R ₅Be hydrogen; And R ₆And R ₇-work forming key.

Or its free alkali;

X wherein ₁ ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate. R ₄Be hydrogen; R ₅Be hydrogen; And R ₆And R ₇Form key together.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₄Be hydrogen; R ₅Be hydrogen; R ₆And R ₇Form key together; And R ₂₇Be hydrogen.

Or its free alkali;

X wherein ₁ ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate. R ₄Be hydrogen; R ₅Be hydrogen; R ₆And R ₇Form key together; And R ₂₇Be hydrogen.

X wherein ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, cyclamate, thiocyanate-, naphthalene-2-sulfonic acid root and oxalate.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein X ^-Be chlorine.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein X ^-It is bromine.

In one embodiment, the present invention relates to contain any aforesaid compound and amino acid whose compsn.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, wherein amino acid is selected from

In another embodiment, the invention provides formula 4 compounds:

Or its pharmacy acceptable salt, wherein each variable independently is:

W is oxygen or sulphur;

Z is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

N equals 0,1 or 2;

M equals 0,1 or 2;

R ₄Be selected from H, alkyl, aralkyl and formula 4a group:

R ₅And R ₆All be hydrogen; Or R ₅And R ₆Form key together;

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₂₀, R ₂₁, R ₂₂, R ₂₃, R ₂₄, R ₂₅It is methyl; R ₂₆Be hydrogen; Q is a key; And Z and W are oxygen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₃Be hydrogen, alkyl, alkenyl, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₄Be hydrogen or have formula 1a structure:

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₅And R ₆Form key together.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein X and Y are-CH ₂-.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein n equals 0; And m equals 0 or 1.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈And R ₂Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen; And R ₃Be hydrogen, alkyl, alkenyl, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁Be hydroxyl or-OC (O) R ₈R ₂Be hydrogen; R ₃Be hydrogen, alkyl, alkenyl, naphthenic base, aralkyl, heteroaralkyl or-[(CR ₂) _p]-R ₁₆And R ₄Be hydrogen or have formula 1a structure:

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical; And R ₅And R ₆Form key together.

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical; R ₅And R ₆Form key together; And X and Y are-CH ₂-.

R wherein ₁₇Independently be selected from hydrogen, halogen, hydroxyl, alkoxyl group, aryloxy, acyloxy, amino, alkylamino, arylamino, acyl amino, aryl alkyl amino, nitro, sulfo-acyl group, carboxylic acid amides, carboxyl, nitrile ,-COR ₁₈,-CO ₂R ₁₈,-N (R ₁₈) CO ₂R ₁₉,-OC (O) N (R ₁₈) (R ₁₉) ,-N (R ₁₈) SO ₂R ₁₉,-N (R ₁₈) C (O) N (R ₁₈) (R ₁₉) and-CH ₂The O-heterocyclic radical; R ₅And R ₆Form key together; X and Y are-CH ₂-; N equals 0; And m equals 0 or 1.

Wherein each variable independently is:

N equals 0,1 or 2;

M etc. 0,1 or 2

R ₁Be hydroxyl or-OC (O) R ₈

R ₅Or R ₆All be hydrogen; Or R ₅With R ₆Form key together;

P is 1,2,3,4,5 or 6;

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; And R ₃It is allyl group.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; And R ₅And R ₆Form key together.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₅And R ₆Form key together; And R ₂₇Be hydrogen.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₅And R ₆Form key together; R ₂₇Be hydrogen; And X and Y are-CH ₂-.

In certain embodiments, the present invention relates to aforesaid compound and subsidiary definition, wherein R ₁It is hydroxyl; R ₃It is allyl group; R ₅And R ₆Form key together; R ₂₇Be hydrogen; X and Y are-CH ₂-; N equals 0; And m equals 0 or 1.

In one embodiment, the present invention provides and is selected from following compound:

and?

Above-mentioned embodiment and following chapters and sections comprise the quinhydrones analogue of NSC 122750 family molecule.Except 17-AAG reduction form (17-allyl amino-18,21-dihydro-17-demethoxylation NSC 122750); Other preferred The compounds of this invention relates to 18; 21-dihydro-NSC 122750 family; Include but not limited to 17-amino-4,18 of 5-dihydro-17-demethoxylation-NSC 122750,21-dihydro analogue; 17-methylamino-4,5-dihydro-17-demethoxylation NSC 122750; 17-cyclopropyl amino-4,5-dihydro-17-demethoxylation NSC 122750; 17-(2 '-hydroxyethyl is amino)-4,5-dihydro-17-demethoxylation NSC 122750; 17-(the 2-methoxy ethyl is amino)-4,5-dihydro-17-demethoxylation NSC 122750; 17-(2 '-fluoro ethyl is amino)-4,5-dihydro-17-demethoxylation NSC 122750; 17-(S)-(+)-2-hydroxypropyl amino-4,5-dihydro-17-demethoxylation NSC 122750; 17-azetidine-1-base-4,5-dihydro-17-demethoxylation NSC 122750; 17-(3-hydroxy azetidine-1-yl)-4,5-dihydro-17-demethoxylation NSC 122750; 17-azetidine-1-base-4,5-dihydro-11-α-fluoro-17-demethoxylation NSC 122750; 17-(2 '-cyano ethyl is amino)-17-demethoxylation NSC 122750; 17-(2 '-fluoro ethyl is amino)-17-demethoxylation NSC 122750; 17-amino-22-(2 '-the methoxybenzoyl methyl)-17-demethoxylation NSC 122750; 17-amino-22-(3 '-the methoxybenzoyl methyl)-17-demethoxylation NSC 122750; 17-amino-22-(4 '-the chlorobenzoyl methyl)-17-demethoxylation NSC 122750; 17-amino-22-(3 ', 4 '-the dichloro-benzoyl methyl)-17-demethoxylation NSC 122750; 17-amino-22-(4 '-amino-3 '-iodobenzene formyl methyl)-17-demethoxylation NSC 122750; 17-amino-22-(4 '-azido--3 '-iodobenzene formyl methyl)-17-demethoxylation NSC 122750; 17-amino-11-α-fluoro-17-demethoxylation NSC 122750; 17-allyl amino-11-α-fluoro-17-demethoxylation NSC 122750; 17-propargyl amino-11-α-fluoro-17-demethoxylation NSC 122750; 17-(2 '-fluoro ethyl is amino)-11-α-fluoro-17-demethoxylation NSC 122750; 17-azetidine-1-base-11-(4 '-the azido-phenyl) sulphonamide carbonyl-17-demethoxylation NSC 122750; 17-(2 '-fluoro ethyl is amino)-11-ketone group-17-demethoxylation NSC 122750; 17-azetidine-1-base-11-ketone group-17-demethoxylation NSC 122750; And 17-(3 '-hydroxy azetidine-1-yl)-11-ketone group-17-demethoxylation NSC 122750.

Art technology person will be interpreted as that the method that this paper lists can be used in the substituted benzoquinones Ansamycin of any amino.

Compsn of the present invention exists as reductive Ansamycin salt, like HCl or H ₂SO ₄Salt.In another embodiment, this compound and another kind of salt are such as amino acid such as glycocoll cocrystallization.Usually, in these embodiments, the ratio of amino acid and Ansamycin can change, but preferred 2: 1-1: 2 amino acid: Ansamycin.

Compsn and preparation

The present invention also provides the compsn that contains any aforesaid compound and at least a pharmaceutically acceptable carrier.

In one embodiment, the invention provides a kind of pharmaceutical composition, contain: at least a pharmaceutically acceptable vehicle; With formula 6 compounds:

Or its free alkali;

Wherein each variable independently is selected from:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X is the conjugate base of pharmaceutical acceptable acid;

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

-N (R ₁₈) COR ₁₉,-N (R ₁₈) C (O) OR ₁₉,-N (R ₁₈) SO ₂(R ₁₉) ,-CON (R ₁₈) (R ₁₉) ,-OC (O) N (R ₁₈) (R ₁₉) ,-SO ₂N (R ₁₈) (R ₁₉) ,-N (R ₁₈) (R ₁₉) ,-OC (O) OR ₁₈,-COOR ₁₈,-C (O) N (OH) ((R ₁₈) ,-OS (O) ₂OR ₁₈,-S (O) ₂OR ₁₈,-OP (O) (OR ₁₈) (OR ₁₉) ,-N (R ₁₈) P (O) (OR ₁₈) (OR ₁₉) and-P (O) (OR ₁₈) (OR ₁₉);

P is 1,2,3,4,5 or 6;

R for each time appearance ₂₀, R ₂₁, R ₂₂, R ₂₄And R ₂₅It independently is alkyl; R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

In certain embodiments, the present invention relates to foregoing and subsidiary definition, condition is to work as R ₁Be hydroxyl, R ₂Be hydrogen, R ₅Be hydrogen, R ₆And R ₇Form two keys together, R ₂₀Be methyl, R ₂₁Be methyl, R ₂₂Be methyl, R ₂₃Be methyl, R ₂₄Be methyl, R ₂₅Be methyl, R ₂₆Be hydrogen, R ₂₇Be hydrogen, Q is that key and W are when being oxygen; R ₃And R ₄Not hydrogen entirely, also do not represent unsubstituted azetidine when combining.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain inhibitor.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain buffer reagent.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain metal chelator.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain inhibitor and buffer reagent.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain inhibitor and metal chelator.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain buffer reagent and metal chelator.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain inhibitor, buffer reagent and metal chelator.

In certain embodiments; The present invention relates to foregoing and subsidiary definition; Wherein said oxidation inhibitor is xitix, halfcystine hydrogen chlorate, sodium sulfite anhy 96, sodium metabisulfite, S-WAT, thioglycerin, Thioglycolic acid sodium salt, sodium formaldehyde sulphoxylate, anti-bad blood acyl cetylate, butylated hydroxyanisole,

DBPC

2,6 ditertiary butyl p cresol, Yelkin TTS, Tenox PG or alpha-tocopherol.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, wherein said oxidation inhibitor is xitix.

In certain embodiments; The present invention relates to foregoing and subsidiary definition, wherein said buffer reagent is Citrate trianion, ascorbate salt, phosphoric acid salt, hydrocarbonate, carbonate, fumarate, acetate, tartrate, malate, SUMATRIPTAN SUCCINATE, lactic acid salt, maleic acid salt, glycocoll or other naturally occurring α or beta-amino acids.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, wherein said buffer reagent is a Citrate trianion.

In certain embodiments; The present invention relates to foregoing and subsidiary definition, wherein said metal chelator is Hydrocerol A, YD 30 (EDTA) and salt thereof, DTPA (diethylene base-triamine-five-acetate) and salt, EGTA and salt thereof, NTA (nitrilotriacetic acid) and salt, sorbyl alcohol and salt thereof, tartrate and salt thereof, N-hydroxyiminodiacetic acid and salt, hydroxyethyl-ethylenediamino tetraacetic acid and salt thereof, 1-and 3-trimethylenedinitrilo-tertraacetic acid and their salt, 1-and 3-diamino--2-hydroxy propane four-acetate and their salt, gluconic acid sodium salt, hydroxyl ethane di 2 ethylhexyl phosphonic acid and salt thereof or phosphoric acid and salt thereof.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, wherein said metal chelator is EDTA.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, wherein said buffer reagent is a Citrate trianion, and said inhibitor is an xitix, and said metal chelator is EDTA.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.001 to about 0.1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.01 to about 0.05 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said xitix and said formula 6 compounds about 0.001 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said xitix and said formula 6 compounds about 0.01 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said Citrate trianion and said formula 6 compounds about 0.05 to about 2 scopes.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said Citrate trianion and said formula 6 compounds about 0.2 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.001 to about 0.1 scope; And the mol ratio of said xitix and said formula 6 compounds about 0.001 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.01 to about 0.05 scope; And the mol ratio of said xitix and said formula 6 compounds about 0.01 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.001 to about 0.1 scope; The mol ratio of said xitix and said formula 6 compounds about 0.001 to about 1 scope; And the mol ratio of said Citrate trianion and said formula 6 compounds about 0.05 to about 2 scopes.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.01 to about 0.05 scope; The mol ratio of said xitix and said formula 6 compounds about 0.01 to about 1 scope; And the mol ratio of said Citrate trianion and said formula 6 compounds about 0.2 to about 1 scope.

In certain embodiments, the present invention relates to the pharmaceutical composition of aforementioned any compsn, further contain solubilizing agent.

In certain embodiments; The present invention relates to foregoing and subsidiary definition, wherein said solubilizing agent is polyoxyethylene glycol sorb polyalcohols fatty ester, polyethylene glycol stearate, benzylalcohol, ethanol, polyoxyethylene glycol, Ucar 35, glycerine, Schardinger dextrins or Prist.

In one embodiment, the invention provides a kind of pharmaceutical composition, contain: at least a pharmaceutically acceptable vehicle; Formula 6 compounds:

Or its free alkali; With formula 10 compounds or its pharmacy acceptable salt, wherein said formula 10 compounds exist to about 5% (m/v) scope with about 0.00001%:

Wherein each variable independently is selected from:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

The absolute stereo chemistry of

formula

DBPC

2,6 ditertiary butyl p cresol, Yelkin TTS, Tenox PG or alpha-tocopherol.

In certain embodiments; The present invention relates to foregoing and subsidiary definition, wherein said metal chelator is Hydrocerol A, YD 30 (EDTA) and salt thereof, DTPA (diethylene base-triamine-five-acetate) and salt, EGTA and salt thereof, NTA (nitrilotriacetic acid) and salt, sorbyl alcohol and salt thereof, tartrate and salt thereof, N-hydroxyiminodiacetic acid and salt, hydroxyethyl-YD 30 and salt thereof, 1-and 3-trimethylenedinitrilo-tertraacetic acid and their salt, 1-and 3-diamino--2-hydroxy propane four-acetate and their salt, gluconic acid sodium salt, hydroxyl ethane di 2 ethylhexyl phosphonic acid and salt thereof or phosphoric acid and salt thereof.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said Hydrocerol A and said formula 6 compounds about 0.05 to about 2 scopes.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said Hydrocerol A and said formula 6 compounds about 0.2 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.001 to about 0.1 scope; The mol ratio of said xitix and said formula 6 compounds about 0.001 to about 1 scope; And the mol ratio of said Hydrocerol A and said formula 6 compounds about 0.05 to about 2 scopes.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, the mol ratio of wherein said EDTA and said formula 6 compounds about 0.01 to about 0.05 scope; The mol ratio of said xitix and said formula 6 compounds about 0.01 to about 1 scope; And the mol ratio of said Hydrocerol A and said formula 6 compounds about 0.2 to about 1 scope.

In certain embodiments, the present invention relates to foregoing and subsidiary definition, further contain solubilizing agent.

In certain embodiments, the present invention relates to the pharmaceutical composition of arbitrary foregoing, wherein said formula 6 compounds exist to the concentration of about 0.160M with about 0.00016M.

In certain embodiments, the present invention relates to the pharmaceutical composition of arbitrary foregoing, wherein said formula 6 compounds exist with about concentration of 0.00032 to about 0.080M.

The preparation method

Can adopt many methods to produce The compounds of this invention.Usually, these steps comprise that (1) is converted into the amino analogue of 17-demethoxylation-17-(like, 17-AAG) with Ansamycin, and the benzoquinones in (2) reduction Ansamycin generates quinhydrones, and (3) use the said quinhydrones of Bronsted s.t. to obtain The compounds of this invention.

The macrocycle molecule that comprises benzoquinones can obtain (for example referring to WO 03/072794 and USP 3,595,955) via the strain fermentation that produces compound.Alternatively, can generate Ansamycin (referring to USP 5,387,584 and WO00/03737) through synthetic or semisynthesis.In addition, isolating fermented product of commercial offers such as NSC 122750 are arranged; Therefore, these materials are easy to obtain.

In preferred embodiments, use currently known methods, use its analogue of synthesis method preparation from organism separating natural product.For example, NSC 122750 separates from the fermenting culture of suitable microbe body and can use many functionalization reactions known in the art to derive.Representational instance comprises the coupled reaction, oxidizing reaction, reduction reaction of metal catalytic, with the nucleophilic reagent reaction, with electrophilic reagent reaction, pericyclic reaction, assembling protection base, remove the protection base, or the like.Many methods of the similar thing of the multiple benzoquinones Ansamycin of generation known in the art (for example, referring to U.S. Patent number 4,261,989,5,387,584 and 5,932,566 and J Med.Chem.1995,38,3806-3812, this paper draw be with reference to).It is of the present invention 18 that the method reduction that these analogues are easy to list through following obtains, the 21-dihydro derivative.

In case the acquisition initiator, with benzoquinones be reduced to quinhydrones then with acid for example HCl reaction generate C-17 ammonium quinhydrones Ansamycin, it is a salt form stable in the air.In the embodiment that can supply replace, quinhydrones free alkali and amino acid carboxylic acid halides but not Bronsted acid-respons, the C-17 ammonium quinhydrones Ansamycin that generates air-stable is salt derivative altogether.This method is in embodiment 3 illustrated.

Many methods and reaction conditions can be used for reducing the benzoquinones part of Ansamycin.Sodium sulfite anhy 96 can be used as reductive agent.Other reductive agent of available includes but not limited to zinc powder and diacetyl oxide or acetate, xitix and electrochemical reduction.

The reduction of Ansamycin verivate benzoquinones part can realize through in two-phase reaction mixture, using sodium sulfite anhy 96.Usually, NSC 122750 is dissolved among organic solvent such as the EtOAc.Other solvent of available includes but not limited to methylene dichloride, chloroform, ethylene dichloride, chlorobenzene, THF, MeTHF, ether, diglyme, 1,2-glycol dimethyl ether, MTBE, THP, dioxane, 2-ethoxy butane, methyl butyl ether, methyl acetate, 2-butanone, water and composition thereof.The aqueous solution (5-30% (m/v), preferred 10% (m/v)) that at room temperature in reaction vessel, adds two equivalents or more sodium sulfite anhy 96s then.Aqueous solution of sodium bisulfite is unsettled and needs just prepared fresh before using thus.This two-phase mixture of vigorous stirring is to guarantee rational speed of response.

This can be reflected at this step and can observe easily through vision, because starting raw material 17-AAG is a purple, along with reaction process produces dihydro-17AAG, purple disappears, and presents yellow.But HPLC/UV or other analytical procedure also can be used for this reaction of monitoring.

When reaction was accomplished, the crude reaction mix products can be used for that next step need not purifying so that the quinhydrones oxidation minimum.But, if the monitoring condition then can be carried out purifying to keep the reduction form of benzoquinones, preferred recrystallization.

The Ansamycin that comprises quinhydrones is unstable, and in the presence of small amount of oxygen or other oxygenant, the quinhydrones part can be oxidized to quinone fast.Apparently, quinhydrones is through being converted into the air-stable thing with acid-respons or with the reaction of amino acid carboxylic acid halides.In an embodiment, the C-17 allyl amino by protonated to generate the C-17 ammonium salt quinhydrones NSC 122750 analogue of multiple air-stable.In addition, formed C-17 ammonium salt quinhydrones has new advantage, promptly the high-dissolvability in the aqueous solution (＞200mg/mL), be different from 17-AAG (＜100 μ g/mL).

The ammonium salt quinhydrones forms through under nitrogen, acid as the solution adding of HCl in organic solvent such as EtOAc, DCM, IPA or dioxane being comprised in the organic solution of Ansamycin of quinhydrones; Organic solvent can independently be acetone, methylene dichloride, chloroform, ethylene dichloride, chlorobenzene, THF, MeTHF, ether, diglyme, 1,2-glycol dimethyl ether, MTBE, THP, dioxane, 2-ethoxy butane, methyl butyl ether, methyl acetate, 2-butanone.

When wherein product precipitated from solution, the ammonium salt of quinhydrones was collected through filtering.When the ammonium salt of quinhydrones did not precipitate, the concentrating under reduced pressure reaction soln obtained product.

The quinhydrones Ansamycin ammonium salt of many air-stables can be through using organic or inorganic acid synthetic.Spendable acid includes but not limited to HCl, HBr, H ₂SO ₄, methylsulfonic acid, Phenylsulfonic acid, tosic acid, trifluoromethanesulfonic acid, camphorsulfonic acid, naphthalene-1,5-disulfonic acid, ethane-1,2-disulfonic acid, Cyclamic Acid, thiocyanic acid, naphthalene-2-sulfonic acid, oxalic acid, or the like.For example referring to Berge et al. (1977) " Pharmaceutical Salts ", J.Pharm.Sci.66:1-19.The pKa of the preferred acid of using should be enough to make that aniline nitrogen is protonated.Thus, pKa approximately-10 to about 7, preferred approximately-10 to about 4, more preferably from about-10 to about 1, be more preferably approximately-10 and can be used for producing the quinhydrones ammonium salt to any acid of-3 approximately.

The present invention further provides the method for recrystallization The compounds of this invention.In these methods, through dissolved compound in minimum inert polar organic solvent such as MeOH, EtOH or IPA, and slowly add miscible organic solvent such as aliphatic ether, ETHYLE ACETATE, methyl acetate, chloroform or DCM, make that solution is muddy and realize.Allow the standing mixt appropriate time then, and optional cooling, the gained solid collected, washing and drying under reduced pressure through filtering.

One aspect of the present invention relates to the preparation method of compound, comprising: hybrid 7 compounds and reductive agent production 8 compounds in reaction solvent; And

Mix said formula 8 compounds and said formula 1 compound of pharmaceutically acceptable acid generation;

Wherein each variable independently is:

W is oxygen or sulphur;

Q is oxygen, NR, N (acyl group) or key;

X ^-It is the conjugate base of pharmaceutical acceptable acid;

R ₅Be selected from H, alkyl, aralkyl and formula 1a group:

R ₆And R ₇All be hydrogen; Or R ₆And R ₇Form key together;

P is 1,2,3,4,5 or 6;

R ₂₃Be alkyl ,-CH ₂OH ,-CHO ,-COOR ₁₈Or-CH (OR ₁₈) ₂

In one embodiment, the present invention relates to preceding method, wherein said reductive agent is sodium sulfite anhy 96, zinc, xitix or electrochemical reduction.

In one embodiment, the present invention relates to preceding method, wherein said reductive agent is a sodium sulfite anhy 96.

In one embodiment; The present invention relates to preceding method; Wherein said reaction solvent is methylene dichloride, chloroform, ethylene dichloride, chlorinated benzene, THF, 2-methyl THF, ether, diglyme, 1,2-glycol dimethyl ether, MTBE, THP, dioxane, 2-ethoxy butane, methyl butyl ether, ETHYLE ACETATE, methyl acetate, 2-butanone, water or its mixture.

In one embodiment, the present invention relates to preceding method, wherein said reaction solvent is the mixture of ETHYLE ACETATE and water.

In one embodiment, the present invention relates to preceding method, the pKa of wherein said acid in water is approximately between-10 to about 7.

In one embodiment, the present invention relates to preceding method, the pKa of wherein said acid in water is approximately between-10 to about 4.

In one embodiment, the present invention relates to preceding method, the pKa of wherein said acid in water is approximately between-10 to about 1.

In one embodiment, the present invention relates to preceding method, the pKa of wherein said acid in water is approximately between-10 to about-3.

In one embodiment, the present invention relates to preceding method, wherein said acid is HCl, HBr, H ₂SO ₄, methylsulfonic acid, Phenylsulfonic acid, tosic acid, trifluoromethanesulfonic acid, camphorsulfonic acid, naphthalene-1,5-disulfonic acid, ethane-1,2-disulfonic acid, Cyclamic Acid, thiocyanic acid, naphthalene-2-sulfonic acid or oxalic acid.

In one embodiment, the present invention relates to preceding method, wherein said acid is HCl.

In one embodiment, the present invention relates to preceding method, wherein said acid is HBr.

In one embodiment, the present invention relates to preceding method, wherein said acid adds as gas.

In one embodiment, the present invention relates to preceding method, wherein said acid is dissolved in the organic solvent.

In one embodiment; The present invention relates to preceding method; Wherein said organic solvent is EtOAc, DCM, IPA or dioxane; Relate to the organic solution that comprises the quinhydrones Ansamycin such as at acetone, methylene dichloride, chloroform, ethylene dichloride, chlorinated benzene, THF, 2-methyl THF, ether, diglyme, 1, the solution in 2-glycol dimethyl ether, MTBE, THP, dioxane, 2-ethoxy butane, methyl butyl ether, methyl acetate or the 2-butanone.

In one embodiment, the present invention relates to preceding method, wherein R ₁It is hydroxyl; R ₂Be hydrogen; R ₃It is allyl group; R ₄Be hydrogen; R ₅Be H; R ₆And R ₇Form key together; R ₂₀It is methyl; R ₂₁It is methyl; R ₂₂It is methyl; R ₂₃It is methyl; R ₂₄It is methyl; R ₂₅It is methyl; R ₂₆Be hydrogen; R ₂₇Be hydrogen; W is an oxygen; And Q is a key.

Pharmaceutical composition

When

formula

1 and 3 compounds and their pharmacy acceptable salts during as antiproliferative such as anticarcinogen, they can be united separately or with pharmaceutically acceptable carrier or thinner according to the pharmacy practice of routine and in pharmaceutical composition, give mammalian subject.These compound taking orally or parenteral admins, preferred parenteral.Parenteral admin comprises intravenously, intramuscular, intraperitoneal, subcutaneous and topical, and preferable methods is an intravenous administration.

Therefore, the invention provides pharmaceutically acceptable compsn, it contains one or more above-claimed cpds (formula 1 and 3) of treating significant quantity, and is formulated together with one or more pharmaceutically acceptable carriers (additive) and/or thinner.Pharmaceutical composition of the present invention can specifically be formulated as solid or liquid form administration, comprises being suitable for following formulation: (1) is as for example sterile solution or suspension-s or sustained release preparation parenteral admin, for example through subcutaneous, intramuscular, intravenously or epidural injection; And (2) oral administration, for example, to irritate and to take medicine (drenches) (moisture or non-aqueous solution or suspension-s), tablet for example purpose is the tablet that is used for oral cavity, hypogloeeis and systemic absorption, pill, pulvis, granula is applied to the paste of tongue.The preferred method that gives The compounds of this invention is parenteral admin (intravenously).

As stated, certain embodiments of the invention can comprise basic functionality such as amino or alkylamino, and can form pharmacy acceptable salt with pharmaceutically acceptable acid thus.Term " pharmacy acceptable salt " is meant the nontoxic relatively inorganic or organic acid addition salt of The compounds of this invention in this regard.These salt can be in the administration vehicle or in the formulation manufacturing processed preparation on the spot, or the free alkali form through pure compound of the present invention and suitable organic or inorganic acid reaction and separate the salt that forms thus in the purifying phase subsequently and prepare separately.Representational salt comprises hydrobromate, hydrogen chlorate, vitriol, hydrosulfate, nitrate salt, acetate, valerate, oleate, palmitate, stearate, lauroleate, benzoate, lactic acid salt, phosphoric acid salt, tosylate, Citrate trianion, PHENRAMINE MALEATE, fumarate, SUMATRIPTAN SUCCINATE, Tartaric acid salt, napthylate, mesylate, gluconate, Lactobionate and lauryl sulfonate or the like.(for example referring to Berge etal. (1977) " Pharmaceutical Salts ", J.Pharm.Sci.66:1-19)

The The compounds of this invention pharmacy acceptable salt comprises the conventional non-toxic salt or the quaternary ammonium salt of compound, for example from non-toxic organic or mineral acid.For example, these conventional non-toxic salt comprise the salt from mineral acid such as spirit of salt, Hydrogen bromide, sulfuric acid, thionamic acid, phosphoric acid, nitric acid etc.; Salt from organic acid such as acetate, propionic acid, succsinic acid, glycol acid, Triple Pressed Stearic Acid, lactic acid, oxysuccinic acid, tartrate, Hydrocerol A, xitix, palmitinic acid, toxilic acid, hydroxymaleic acid, toluylic acid, L-glutamic acid, phenylformic acid, Whitfield's ointment, Sulphanilic Acid, 2-acetoxy-benzoic acid, fumaric acid, tosic acid, methylsulfonic acid, ethane disulfonic acid, oxalic acid, different thionic acid etc.

In other cases, compound of the present invention can comprise one or more acidic functionalities, and can form pharmaceutically-acceptable salts with pharmaceutically acceptable alkali thus.Term " pharmacy acceptable salt " is meant nontoxic relatively, the inorganic and organic bases additive salt of The compounds of this invention in these cases.These salt can prepare in the process of administration vehicle or manufacturing formulation equally on the spot; The free acid form through pure compound and suitable alkali and ammonia or with pharmaceutically acceptable organic primary, second month in a season or tertiary amine separately reaction prepare, the alkali that suits is such as oxyhydroxide, carbonate or the supercarbonate of pharmaceutically acceptable metallic cation.Representational alkali or alkaline earth salt comprise lithium, sodium, potassium, calcium, magnesium and aluminium salt or the like.Be applicable to that the representational organic amine that forms base addition salt comprises ethamine, diethylamine, quadrol, thanomin, diethylolamine, piperazine, or the like.(for example referring to Berge et al., as above)

Wetting agent, emulsifying agent and lubricant also may reside in the said composition such as Sodium Lauryl Sulphate BP/USP and Magnesium Stearate and tinting material, releasing agent, coating-forming agent, sweeting agent, seasonings and spices, sanitas, solubilizing agent, buffer reagent and inhibitor.

Pharmaceutically acceptable inhibitor is for example including, but not limited to (1) water soluble antioxidant, such as xitix, cysteine hydrochloride, sodium sulfite anhy 96, Sodium Pyrosulfite, S-WAT, thioglycerin, Thioglycolic acid sodium salt and sodium formaldehyde sulphoxylate; (2) oil-soluble inhibitor is such as anti-bad blood acyl cetylate, butylated hydroxyanisole (BHA),

DBPC

2,6 ditertiary butyl p cresol (BHT), Yelkin TTS, propyl gallate, alpha-tocopherol.

Pharmaceutically acceptable buffer reagent is for example including, but not limited to Citrate trianion, xitix, phosphoric acid salt, supercarbonate, carbonate, fumarate, acetate, tartrate and malate.

The pharmaceutically acceptable solubilizing agent is for example including, but not limited to polyoxyethylene sorbitan fatty acid ester (comprising polysorbate 80), polyoxyethylene stearic acid ester, benzylalcohol, ethanol, polyoxyethylene glycol, Ucar 35, glycerine, Schardinger dextrins and Prist.

Pharmaceutically acceptable complexing agent for example including, but not limited to, Schardinger dextrins (α, beta, gamma), especially substituted beta Schardinger dextrins are such as 2-hydroxypropyl-β, dimethyl beta, 2-hydroxyethyl β, 3-hydroxypropyl β, trimethylammonium β.

Pharmaceutically acceptable metal chelator for example including, but not limited to; Hydrocerol A, diethylamine tetraacethyl (EDTA) and salt thereof, DTPA (diethylene triaminepentaacetic acid(DTPA)) and salt thereof, EGTA and salt thereof, NTA (nitrilotriacetic acid) and salt thereof, Sorbitol Powder and and salt, tartrate and and salt, N-hydroxyl imide diacetate esters and salt, hydroxyethyl-ethylenediamino tetraacetic acid and salt thereof, 1-and 3-trimethylenedinitrilo-tertraacetic acid and salt, 1-and 3-diamino--2-hydroxy propane four-acetate and salt, gluconic acid sodium salt, hydroxyl ethane di-phosphate and salt thereof and phosphoric acid and salt thereof.

Prepare these preparations or method for compositions and comprise the step that makes that The compounds of this invention combines with carrier and one or more optional auxiliary agents.Usually; Make The compounds of this invention evenly combine (liquid preparation) closely when preparing these preparations with liquid vehicle; Evenly combine closely with liquid vehicle, freeze-drying subsequently handles (with reconstituted pulvis such as sterilized waters) or all has with the solid carrier of fine powder or the two and evenly combine nearly; Subsequently if necessary, be shaped or pack this product.

The medicine present composition that is suitable for administered parenterally contains one or more The compounds of this invention or its pharmaceutically-acceptable salts and has united one or more pharmaceutically acceptable sterile isotonics moisture or non-aqueous solution, dispersion liquid, suspension-s or emulsion or sterile powder; It can reformulate sterile injectable solution or dispersion liquid before just will use, and it can comprise sugar, alcohol, inhibitor, buffer reagent, fungistat, sequestrant, make preparation and the isoosmotic solute of intended recipient's blood, suspension agent or thickening material.In an embodiment, activeconstituents combines in solution with pharmaceutically acceptable carrier, and lyophilize is to obtain dry powder then.Dry powder package is reconstituted original formulation through adding sterile solution to this pulvis such as water or saline water then in unit dosage.

The suitable moisture and nonaqueous carrier of pharmaceutical composition of the present invention for example comprises water, ethanol, polyvalent alcohol (such as glycerine, Ucar 35, polyoxyethylene glycol, or the like) and suitable mixture, vegetables oil is such as sweet oil, and injectable organic ester is such as OE.For example, for dispersion agent, keep required particle diameter, and use tensio-active agent, can keep suitable flowability through using coating materials such as Yelkin TTS.

These compsns also can comprise auxiliary agent such as sanitas, wetting agent, emulsifying agent and dispersion agent.Can ensure through containing various antibiotic and anti-mycotic agents such as p-hydroxybenzoic acid, butylene-chlorohydrin, phenol Sorbic Acid etc. for the effect of The compounds of this invention prophylaxis of microbial.Also expect in the said composition to comprise isotonic agent such as sugar, sodium-chlor, or the like.In addition, can cause the absorption of injectable drug formulation to prolong through comprising the reagent that postpones to absorb such as aluminum monostearate and gelatin.

In some cases, for the effect of prolong drug, expectation delays the absorption of subcutaneous injection or intramuscular injection medicine.This can realize through the crystal of use poorly water-soluble or the liquid suspension of amorphous substance.The absorption rate of medicine depends on its dissolution rate, and dissolution rate depends on crystallographic dimension and crystal formation again.Alternatively, dissolving or suspended drug also can postpone the absorption of parenteral admin medicine in oil medium.

As used herein; Words and expressions " parenteral admin " and " at parenteral admin " be meant in intestines with topical administering mode; Usually through injection; Include but not limited to, in the intravenously, intramuscular, intra-arterial, sheath, in the capsule, interior, intracardiac, the intracutaneous of socket of the eye, intraperitoneal, under tracheae, subcutaneous, epidermis, under the intraarticular, capsule, under the arachnoid membrane, in the backbone and breastbone inner injection and transfusion.

As used herein; " whole body administration ", " administration capapie " and " peripherally administered " and " periphery ground administration " are meant and except direct entering cns, give compound, medicine or other material; Thus; It gets into patient's whole body and carries out metabolism and other similar process, for example subcutaneous injection administration thus.

A kind of preferred formulation of the present invention is to comprise Hydrocerol A (the about 250mM of about 5mM-; The preferred about 150mM of about 25mM-), xitix (the about 250mM of about 0.1mM-; The preferred about 50mM of about 0.1mM-) and edetate (Ca-EDTA-disodium; EDTA, the about 3mM of the preferably about 1mM-of the about 20mM of about 0.2mM-), pH is adjusted to about 3.1 water-containing buffering liquid through sodium hydroxide.These compositions of preparation are respectively as buffer reagent, inhibitor and metal chelator.

Preparation for The compounds of this invention is important for this quinhydrones salt provides solubleness and oxidation-reduction stability.When amine when protonated, The compounds of this invention is at the low obvious solubilising of pH value.The distribution of multi-form compound is important, because the ionization form is more solvable, and the less dissolving of free alkali (unionized form).Therefore, the pH through control solution will make preparation solubleness optimizing.Buffer reagent is a preferred ingredient of said preparation such as the Citrate trianion that in preferred pH scope, has high buffering capacity.

Preferred reducing agents will make preparation about 5.0 at the about 1.5-of pH, and more preferably the about 1.8-of pH is about 3.5, is more preferably about 3.3 bufferings of the about 3-of pH.

Can oxidation when placing for a long time in the quinhydrones analogue solution of the present invention.Heavy metal, such as iron and copper, can the concurrent present general agents of catalytic oxidation and labware in have trace.

The character that prevents the heavy metal oxidation provides such as EDTA (YD 30) through metal chelator.Other known sequestrant such as be Hydrocerol A, DTPA (diethylene base-triamine-pentaacetic acid) and salt, EGTA and salt thereof, NTA (nitrilotriacetic acid) and salt thereof, Sorbitol Powder and and salt, tartrate and and salt, N-hydroxyl imide diacetate esters and salt, hydroxyethyl-ethylenediamino tetraacetic acid and salt thereof, 1-and 3-trimethylenedinitrilo-tertraacetic acid and salt, 1-and 3-diamino--2-hydroxy propane four-acetate and salt thereof, gluconic acid sodium salt, hydroxyl ethane di-phosphate and salt thereof and phosphoric acid and salt thereof.

The important means of another kind of prevention oxidation is to add inhibitor.Preferred anti-oxidants is xitix (vitamins C).These reagent prevent that compound from receiving the oxygenizement of the oxygen molecule of water-soluble medium.In certain embodiments, vitamins C is as the component in the quinhydrones analog formulations of the present invention.

Preparation of the present invention comprises preparation that can subsequent use storage and the preparation that is used for directly giving the patient.Especially, pharmaceutical composition/preparation of the present invention is to provide than the higher form of concentration that is suitable for directly giving the patient.Said composition is diluted into IV bag (IV bag) usually to give the patient.

Importantly, the preparation that is contained in the IV bag was stable at about 5 minutes-Yue 2 hours, was stable at about 1 hour-2 hours more preferably, was stable at about 2 hours most preferably in this is used.Need keep stability in the full stage of administration.

In addition, importantly the surge capability of the IV bag preparation of this dilution is enough to realize its stability, and concentration is too not high in patient's body, causing adverse reaction yet simultaneously.Exist too many buffer reagent can cause many undesirable effect for the patient.

The method of treatment

The present invention provides the compound that comprises water-soluble quinhydrones, and it is oxidized to the amino substituted benzoquinones NSC 122750 analogue (for example 17-AAG) of 17-under physiological pH in vitro and in vivo fast.Thus, quinhydrones analogue of the present invention has shown biological activity and the curative effect that is similar to the amino substituted NSC 122750 analogue of 17-, and can be used for all treatment indications that the amino substituted NSC 122750 analogue of 17-can be treated.17-amino substituted NSC 122750 analogue, particularly 17-AAG are efficiently and the optionally suppressor factor of HSP90.

The present invention further provides treatment, has alleviated disorder or a kind of and multiple symptom of the state of an illness and the method that alleviates its severity that proliferative disorders is cancer and other HSP90 mediation.Because compsn of the present invention is higher than the benzoquinones form solubleness of oxidation, causes using the better clinical effectiveness of arbitrary known parent molecule thereby said composition is easier to administration.

Treat-ment of the present invention relates to the disorder of suffering from HSP90 mediation or the state of an illness The compounds of this invention such as the patient treatment significant quantity of cancer.This paper has described the explanation of compsn, preparation, dosage, administration and therapeutic modality.

Synthesized the amino substituted NSC 122750 analogue of some 17-, their application as antitumour drug is described in U.S. Patent number 4,261; 989 and 5,387,584,5; 932,566 and disclosed PCT application WO 00/03737 and WO 03/072794 (being hereby incorporated by).The structure activity relationship of the amino substituted NSC 122750 analogue of 17-has more clearly illustrated that and has suppressed the required chemical feature of HSP90 (for example referring to J.Med.Chem. (1995) 38:3806-3812, J.Med.Chem. (1995) 38:3813-3820, and Clin.Cancer Res. (1999) 5:3781).

17-AAG is one of amino substituted NSC 122750 analogue of the most successful 17-, and it has widely anti-tumor activity in vitro and in vivo and is used for a plurality of I/II clinical trial phases at present.17-AAG has shown the differential cytotoxicity that resists many tumor types in NCI 60 tumor cell line groups.The average IC of this group all cells system ₅₀Be 120nM (DevelopmentalTherapeutics Program Website:http: //mean chart of dtp.nci.nih.gov/, compound S 330507)

In addition; 17-AAG has showed the activity of resisting many clones, includes but not limited to melanoma (Anti-Cancer Drugs (2004) 15:377-388), prostate cancer (Clin.CancerRes. (2002) 8:986-993), mammary cancer (Cancer.Res. (2001) 61:2945-2952), nonsmall-cell lung cancer (Ann.Thorac.SuYg. (2000) 70:1853-1860), white blood disease (Cancer Res. (2001) 61:1799-1804) and colorectal carcinoma (J.Natl.CancerInst. (2003) 95:1624-1633).

In one embodiment, the present invention provides the treatment method for cancer, comprises arbitrary aforesaid compound of the Mammals treatment significant quantity that needs this treatment, or the arbitrary aforementioned pharmaceutical composition of treatment significant quantity.

In one embodiment, the present invention relates to this aforesaid method, wherein said cancer is the cancer of hemopoietic system, immunity system, endocrine system, pulmonary system, gastro-intestinal system, Musculoskeletal, reproductive system, cns or urinary system.

In one embodiment; The present invention relates to aforesaid method, wherein cancer is arranged in mammiferous myeloid tissue, Lymphoid tissue, pancreatic tissue, parathyroid tissue, lung, colon, rectal tissue, anus tissue, hepatic tissue, skin, bone, ovary tissue, uterine cancer cell, cervical tissue, breast, prostate gland, testis tissue, brain, brain stem, meningitis (meningial) tissue, kidney or bladder.

In one embodiment, the present invention relates to aforesaid method, wherein cancer is arranged in mammiferous myeloid tissue, Lymphoid tissue, breast, lung, ovary or prostate gland.

In one embodiment; The present invention relates to aforesaid method, wherein said cancer is mammary cancer, multiple myeloma, carcinoma of prostate, Hodgkin lymphoma, non-Hodgkin lymphoma, acute lymphoblastic leukemia, lymphocytic leukemia, acute myelogenous leukemia, chronic myelocytic leukemia, renal cell carcinoma, malignant melanoma, carcinoma of the pancreas, lung cancer, colorectal carcinoma, colorectal carcinoma, brain cancer, kidney disease, head and neck cancer, bladder cancer, thyroid carcinoma, carcinoma of prostate, ovary cancer, Cervical cancer or myelodysplastic syndrome.

In one embodiment, the present invention relates to aforesaid method, wherein said mammalian cancer is mammary cancer, acute myelogenous leukemia, chronic myelocytic leukemia, melanoma, multiple myeloma, lung cancer, ovary cancer or carcinoma of prostate.

In one embodiment, the present invention relates to aforesaid method, wherein said Mammals is a primate, horse, dog, cat or ox.

In one embodiment, the present invention relates to aforesaid method, wherein said Mammals is human.

In one embodiment; The present invention relates to aforesaid method, the administering mode of wherein said compound be suck, oral, intravenously, hypogloeeis, eyes, transdermal, rectum, vagina, part, intramuscular, intra-arterial, sheath are interior, subcutaneous, oral cavity or nose administration.

In one embodiment, the present invention relates to aforesaid method, wherein administering mode is intravenous injection.

Conjoint therapy

In another kind of embodiment, the invention provides treat-ment, wherein The compounds of this invention and compsn use parallel connection to be bonded to less a kind of other medicines in the treatment cancer, to realize selective active with ubcellular poison level.In certain embodiments, The compounds of this invention is used to reduce the cell levels of suitably folding HSP90 client protein, suppresses effectively through second kind of medicine then or uses proteasome inhibitor such as Velcade ^TMSuppress its degraded in proteasome.The combining to have stablized this client protein and they are maintained soluble inactivation form of client's albumen and HSP90 stimulates in order to replying to activate.The combining of benzoquinones Ansamycin of the present invention and HSP90 causes client's albumen to concentrate one's gaze on proteasome, and degraded subsequently.Use concentrate one's gaze on and the drug block of arrestin enzyme body the increase of proteasome degraded causing apoptosis and necrocytosis.

Can be used for generally including: alkylating agent with the instance of the inventive method antitumour drug linked together; Anti-angiogenic agent; The antimetabolic product; Epidophylltoxin; Antitumor drug metabolizing enzyme; Topoisomerase enzyme inhibitor; Procarbazine; Mitoxantrone; The platinum coordination complex; Anti--mitogenic agent; BRM and growth inhibitor; Hormone/hormone antagonist medicine and hemopoieticgrowth factor.

The exemplary types of antitumour drug also comprises anthracycline antibiotics family medicine, Vinca medicine, MTC, bleomycin, cell toxicant nucleosides, epothilones, discodermolide, pteridine family medicine, diynenes and podophyllotoxin.

The useful especially member of those types for example comprises: canninomycin, daunorubicin, AMT, methotrexate, methopterin, dichloromethane AMT, ametycin, porfiromycin, 5-; Ismipur, gemcitabine, cytosine arabinoside, podophyllotoxin or podophyllotoxin verivate be such as etoposide, etoposide phosphate radical or teniposide, melphalan, vincaleucoblastine, vincristine(VCR), LEU, velcade, adriamycin, desacetyl vinblastine amide, leurosine, STI571, taxol, taxol, or the like.In preferred embodiments, antineoplastic agent is velcade, adriamycin, TX, Docetaxel, taxol, suitable-court of a feudal ruler, pula, STI571 or gemcitabine.In preferred embodiments, antineoplastic agent is velcade or adriamycin.

Other useful antineoplastic agent comprises Emcyt, carboplatin, endoxan, bleomycin, gemcitabine, ifosamide; Melphalan, hexamethyl trimeric cyanamide, thiotepa, cytarabin, idatrexate, Trimetrexate, dacarbazine, left-handed l-asparagine, NSC 94600, CPT-11, hycamtin, cytarabin, bicalutamide, Drogenil, TAP-144, pyrido benzindole (pyridobenzoindole) verivate, Interferon, rabbit and interleukin-.

Can give chemotherapeutic and/or radiotherapy according to regimen well known in the art.Those skilled in the art are obviously known, depend on that the disease of being treated and chemotherapeutic and/or radiotherapy can change for the known effect of these diseases to give chemotherapeutic and/or radiotherapy.In addition, according to the knowledge of skilled clinician, change regimen (for example, dosage and time) for patient's influence and viewed disease for replying of giving medicine according to viewed administration (that is, antitumour drug or radiation).

In addition, usually, The compounds of this invention and chemotherapeutic needn't give in same pharmaceutical composition, and owing to different physics and chemical propertys, possibly have to through the different approaches administration.For example, of the present inventionization contain thing can intravenous administration producing and to keep good blood level, and chemotherapeutic can be taken orally.In the ken of skilled clinician, can confirm the administering mode and the administration reasonableness of in same pharmaceutical composition (if possible).Can carry out initial administration according to definite scheme known in the art, then based on viewed influence, skilled clinician can change dosage, administering mode and administration time.

The concrete selection of chemotherapeutic or radiating will depend on attending doctor's diagnosis and they are for the judgement of conditions of patients and suitable regimen.

The compounds of this invention and chemotherapeutic and/or radiation can be (for example to give simultaneously; Side by side, basically side by side or in the identical treatment scheme) or give in succession; This depends on character, the patient's of hyperplasia cause of disease the state of an illness and this actual selection and chemotherapeutic The compounds of this invention (that is, in the single therapy scheme) linked together/or radiation.

If The compounds of this invention and chemotherapeutic and/or radiation are not side by side or basically side by side to give, The compounds of this invention can be directed against different tumours and difference with chemotherapeutic and/or the best order of administration of radiating so.Therefore, in some situation, The compounds of this invention is administration at first, then gives chemotherapeutic and/or radiation; In other situation, at first can give chemotherapeutic and/or radiation, then give The compounds of this invention.These alternative administrations can repeat during the single therapy scheme.Order of administration and the number of times that repeats to give each medicine during in its ken, can confirming to treat after the disease that the skilled practitioners evaluation is treated and patient's the state of an illness.For example, can at first give chemotherapeutic and/or radiation, when especially it is cell toxicity medicament, gives The compounds of this invention then and continue treatment, being determined here is advantageously, then gives chemotherapeutic and/or radiation, like that, until accomplishing this regimen.

Therefore, according to practice and knowledge, along with the carrying out of treatment, the working doctor can change the component that scenarios gives (medicine, that is, The compounds of this invention, chemotherapeutic or radiation) according to the needs of individual patient.

Dosage

When The compounds of this invention gave the human or animal as medicine, they self can give or as for example comprising 0.1-99% (more preferably, 10-30%) activeconstituents and unite and have the pharmaceutical composition of pharmaceutically acceptable carrier to give.

For concrete patient, compsn and administering mode, the actual metered of activeconstituents can change so that the amount of activeconstituents can produce effectively that the required treatment of patient is replied nontoxic for the patient in the pharmaceutical composition of the present invention.

Selected dosage level will depend on various factors comprise speed and degree, the treatment of drainage or metabolic speed, the absorption of activity, route of administration, the administration time of the used particular compound of the present invention or its salt, employed particular compound time length, unite other medicines, compound and/or material, age, sex, body weight, the state of an illness of use, the patient's that treats general health and before medical history and the known similar factor of medical field with used particular compound.

Doctor or animal doctor with ordinary skill can easily confirm the significant quantity of required pharmaceutical composition and prescribe.For example, doctor or animal doctor can improve dosage until realizing desired result then gradually to be lower than the pharmaceutical composition that begins to give The compounds of this invention for the level of the curative effect required dosage realizing expecting.

Usually, the appropriate dose of The compounds of this invention is the minimum safe effective dose that this compound can produce curative effect.This effective dose will depend on above-mentioned factor usually.Usually, The compounds of this invention gives the patient's vein ID and will be about the every square meter of the about 1000mg of 10mg-, be administered twice weekly, the every square meter of preferably about 75mg-750mg, is administered twice weekly, is more preferably the every square meter of 100mg-500mg, is administered twice weekly.

Although The compounds of this invention can be individually dosed, preferably this compound is as pharmaceutical prepn (compsn) administration.

The patient who accepts this treatment is its animals of any needs, comprise that primate is human especially, and other Mammals is such as horse, ox, pig and sheep; And common poultry and pet.

One or more other active compounds can be added in the above-mentioned preparation to be provided for the preparation of combination cancer therapy.

Embodiment

Described the present invention now prevailingly, will be more readily understood with reference to following embodiment, comprised that these embodiment only are some aspect and the purposes of embodiment in order to demonstrate the invention, but not be intended to limit the present invention.In addition, amino acid is represented as zwitterionic form and exists, can also be further by protonated and exist as salt.

Embodiment 1

The preparation of the hydroquinone derivatives of the NSC 122750 family molecule of air-stable

Reaction process 1

With formula (1a) (1.0 equivalent) compound dissolution in methylene dichloride (0.02M) and with 10% aqueous solution of sodium bisulfite (1: 1; DCM: the aqueous solution) stirred 30 minutes.Remove organic layer via syringe then, water layer is once more through dichloromethane extraction.The organic solution that merges directly joins in the dichloromethane solution (0.001M) of acyl chlorides (1.0 equivalent) through brine wash then.Stir this reaction mixture 12h, in the impouring dichloromethane solution.And then making water (2.0mL) washing organic layer, the water layer of merging obtains product after freeze-drying.

Embodiment 2

Formula (1a) compound (0.25mmol, 1.0 equivalents) is dissolved in methylene dichloride (3mL) and stirred 30 minutes with 10% aqueous solution of sodium bisulfite (1.5mL).Remove organic layer via syringe then, water layer is once more through dichloromethane extraction.The organic solution that merges is diluted through the EtOAc of 3mL, brine wash, and under reduced pressure further remove residual water and EtOAc (the 3mL solvent is removed in decompression down altogether) through azeotropic drying.Add the solution of acid in organic solvent to this solution.Gained solution is cooled to-5 ℃ of toluene solutions (0.2ml) that also add acid (0.25mmol).Solid is separated out from solution lentamente.Add MT BE (3mL) then, permission gained mixture rises to RT and stirred 50 minutes in this temperature.Solid is collected in vacuum filtration then, and (2 * 3mL) washings, drying under reduced pressure obtains product through MTBE.

Embodiment 3

17-AAG quinhydrones dimethylamino acetate is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (9.1mg, 0.016mmol, 1.0 equivalents) is dissolved in the 1.0mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.0mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of dimethylamino Acetyl Chloride 98Min. hydrogen chlorate (2.5mg, 0.016mmol, 1.0 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 2 after freeze-drying, be white fluffy powder (7.1mg, 0.011mmol, 66% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 4

17-AAG quinhydrones α-An Jiyidingsuan is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (16.7mg, 0.0285mmol, 1.0 equivalents) is dissolved in the 1.5mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.5mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (4.4mg, 0.0314mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 3 after freeze-drying, be white fluffy powder (15.1mg, 0.0224mmol, 79% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 5

17-AAG quinhydrones Beta-alanine is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (16.7mg, 0.0285mmol, 1.0 equivalents) is dissolved in the 1.5mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.5mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer extracts through 0.30mL diamino methane once more.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (4.52mg, 0.0314mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 4 after freeze-drying, be white fluffy powder (12mg, 0.0237mmol, 83% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 6

17-AAG quinhydrones sarcosine is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (15.1mg, 0.0258mmol, 1.0 equivalents) is dissolved in the 1.5mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.5mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (3.7mg, 0.0258mmol, 1.0 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 5 after freeze-drying, be white fluffy powder (15.4mg, 0.0234mmol, 91% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 7

17-AAG quinhydrones piperidine carboxylic acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (16mg, 0.027mmol, 1.0 equivalents) is dissolved in the 1.5mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.5mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.25mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (5.5mg, 0.03mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 6 after freeze-drying, be white fluffy powder (11.4mg, 0.019mmol, 60% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 8

17-AAG quinhydrones glycocoll is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (16.2mg, 0.028mmol, 1.0 equivalents) is dissolved in the 1.5mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (1.5mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.25mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (3.4mg, 0.03mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 7 after freeze-drying, be white fluffy powder (3.1mg, 0.0051mmol, 19% yield, 3: 1 phenol regional isomer intermixture).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 9

17-AAG quinhydrones 2-amino-2-ethyl-butyric acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (48mg, 0.082mmol, 1.0 equivalents) is dissolved in the 4.8mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (4.8mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 1mL dichloromethane extraction.The organic solution that merges directly joins in the 1mL dichloromethane solution of acyl chlorides hydrogen chlorate (16.8mg, 0.09mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 8 after freeze-drying, be white fluffy powder (24.7mg, 0.034mmol, 41% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 10

17-AAG quinhydrones 1-amino-cyclopropane-carboxylic acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (48mg, 0.082mmol, 1.0 equivalents) is dissolved in the 4.8mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (4.8mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 1mL dichloromethane extraction.The organic solution that merges directly joins in the 1mL dichloromethane solution of acyl chlorides hydrogen chlorate (14.1mg, 0.09mmol, 1.1 equivalents) through brine wash (1.0mL) then. and stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 9 after freeze-drying, be white fluffy powder (36.2mg, 0.051mmol, 62% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 11

The 17-AAG hydroquinone carboxylic acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (24mg, 0.041mmol, 1.0 equivalents) is dissolved in the 2.4mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (2.4mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (7.8mg, 0.045mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 10 after freeze-drying, be white fluffy powder (25.8mg, 0.038mmol, 92% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 12

17-AAG quinhydrones 1-amino-Cyclopentane carboxylic acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (48mg, 0.082mmol, 1.0 equivalents) is dissolved in the 4.8mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (4.8mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (17mg, 0.09mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 11 after freeze-drying, be white fluffy powder (34.3mg, 0.049mmol, 60% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 13

17-AAG quinhydrones N-methyl piperidine carboxylic acid is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (21.8mg, 0.038mmol, 1.0 equivalents) is dissolved in the 2mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (2mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (8.1mg, 0.041mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 11 after freeze-drying, be white fluffy powder (15.2mg, 0.0213mmol, 56% yield).This material is at D ₂Warp among the O ¹HNMR also analyzes through LC-MS.

Embodiment 14

17-AAG quinhydrones N, N, N-trimethylacetic acid ammonium is the preparation of salt altogether

17-allyl amino geldanamycin mycin (1) (113mg, 0.19mmol, 1.0 equivalents) is dissolved in the 2mL methylene dichloride and with 10% aqueous solution of sodium bisulfite (2mL) stirs.Deep purple solution becomes yellow after 5 minutes, continues to stir this mixture 25 minutes.Remove organic layer via syringe then, water layer is once more through the 0.30mL dichloromethane extraction.The organic solution that merges directly joins in the 0.20mL dichloromethane solution of acyl chlorides hydrogen chlorate (33mg, 0.21mmol, 1.1 equivalents) through brine wash (1.0mL) then.Stir this reaction mixture 2h, impouring contains in the separating funnel of 3.0mL water.The extraction organic layer is also again through the 2.0mL water washing.The water layer that merges obtains 13 after freeze-drying, be white fluffy powder (78mg, 0.11mmol, 57% yield).This material is at CDC1 ₃/ deuterium is for warp among the DMSO (6: 1) ¹HNMR also analyzes through LC-MS.

Embodiment 15

The 17-AAG hydroquinone derivatives for preparing air-stable by NSC 122750

In the 10mL flask, add NSC 122750 (28) (0.14g, 0.25mmol, 1.0 equivalents), add MeTHF (0.625mL) solution of allyl amine (0.075mL, 1.0mmol, 4 equivalents) then.Heating gained slurry to 40 is ℃ 10 hours in nitrogen.Reaction mixture is chilled to room temperature then, uses the MeTHF dilution of 1.0mL, uses saturated NH ₄C1 solution (1.5mL) and saturated NaCl (1.5mL) washing.Collected organic layer uses the aqueous solution of sodium bisulfite (1mL, 20% (m/m)) of prepared fresh to handle, and vigorous stirring is 45 minutes in nitrogen.Remove the aqueous solution then, organic layer washs through the 1.5mL de aerated water.Using MeTHF azeotropic drying organic solution to remove anhydrates.This reduces pressure then through the MeTHF that adds 2mL and realizes in 70 ℃ of concentrated (about 2mL) gained solution.In ice bath, gained solution is cooled to 0 ℃ and in nitrogen, add alpha-amino group isobutyryl chloride hydrogen chlorate (0.04g, 0.25mmol, 1.0 equivalents) then.3 hours after-filtration of stirred reaction mixture are collected solid and through MeTHF (2 * 2mL) washings.The drying under reduced pressure solid obtains product, is yellow powder (171mg, 0.2425mmol, 97% total recovery).

Embodiment 16

The 17-AAG quinhydrones is the crystallization of salt form altogether

Compound 7 is dissolved among the MeOH of minimum, dropwise adds EtOAc then lentamente until continuing muddiness.Allow this mixture to leave standstill then 14 hours and through solid collected by filtration, through EtOAc washing, drying under reduced pressure.

Embodiment 17

The preparation of the NSC 122750 family molecule hydroquinone derivatives of air-stable

Formula (1) compound (0.450g, 0.768mmol, 1.0 equivalents) is dissolved in the methylene dichloride (50mL) and with 10% aqueous solution of sodium bisulfite (50mL) stirs.Stirred this solution 30 minutes.Collected organic layer is through Na ₂SO ₄Round-bottomed flask is filtered and be transferred to drying.The dioxane solution (4N, 0.211mL, 1.1 equivalents) that in this solution, adds HCl.Allow the gained mixture in nitrogen, to stir 30 minutes.Yellow solid is slowly separated out from solution.This yellow solid recrystallization in MeOH/EtOAc obtains the IPI-504 (15) of 0.386g.

Embodiment 18

Formula (1) compound (0.30g, 0.5mmol, 1.0 equivalents) is dissolved among the MTBE (3mL) and with 20% aqueous solution of sodium bisulfite (2mL) stirs.Stirred this solution 60 minutes.Collected organic layer through brine wash, filters and is transferred to flask of the solid end.This solution is chilled to-5 ℃ and place nitrogen.In this solution, dropwise add H ₂SO ₄The Denatured alcohol (H of 0.50mmol ₂SO ₄In the EtOH of 0.5mL) solution.Allow the gained mixture in nitrogen, to stir and rise to RT.Should concentrate then in 30 minutes by the yellow slurry in the RT restir.Add MTBE (7mL) and filtering suspension liquid.Collect yellow solid, through the MTBE washing, drying under reduced pressure obtains the required product of 0.30g.

Embodiment 19

Formula (1) compound (0.30g, 0.5mmol, 1.0 equivalents) is dissolved among the DCM (6mL) and with 10% aqueous solution of sodium bisulfite (3.5mL) stirs.Stirred this solution 60 minutes.Collected organic layer through brine wash, is transferred to round-bottomed flask with 1.2mL (through being calculated as the quinhydrones of 0.1mmol).This solution is placed nitrogen.Sex change IPA (tosic acid of 0.100mmol is in the IPA of the 0.25mL) solution that in this solution, dropwise adds tosic acid.Allow the gained mixture in nitrogen, to stir 1 hour, enriched mixture at this moment, bullion is severe slurry from EtOAc/MTBE.Obtain the required product of 0.068g through solid collected by filtration and drying under reduced pressure.

Embodiment 20

Formula (1) compound (0.30g, 0.5mmol, 1.0 equivalents) is dissolved among the DCM (6mL) and with 10% aqueous solution of sodium bisulfite (3.5mL) stirs.Stirred this solution 60 minutes.Collected organic layer through brine wash, is transferred to round-bottomed flask with 1.2mL (through being calculated as the quinhydrones of 0.1mmol).Sex change IPA (the d-camphorsulfonic acid of 0.100mmol is in the IPA of the 0.25mL) solution that in this solution, dropwise adds the d-camphorsulfonic acid.Allow the gained mixture in nitrogen, to stir 1 hour, enriched mixture at this moment, bullion is severe slurry from EtOAc/MTBE.Obtain the required product of 0.051g through solid collected by filtration and drying under reduced pressure.

Embodiment 21

Formula (1) compound (0.30g, 0.5mmol, 1.0 equivalents) is dissolved among the DCM (6mL) and with 10% aqueous solution of sodium bisulfite (3.5mL) stirs. stirred this solution 60 minutes.Collected organic layer through brine wash, is transferred to round-bottomed flask with 1.2mL (through being calculated as the quinhydrones of 0.1mmol).In this solution, dropwise add H ₃PO ₄The sex change IPA (H of 0.100mmol ₃PO ₄In the IPA of 0.25mL) solution.Allow the gained mixture in nitrogen, to stir 1 hour, enriched mixture at this moment, bullion is severe slurry from EtOAc/MTBE.Obtain the required product of 0.050g through solid collected by filtration and drying under reduced pressure.

Embodiment 22

Formula (1) compound (0.50g, 0.8mmol, 1.0 equivalents) is dissolved among the DCM (8mL) and with 15% aqueous solution of sodium bisulfite (4mL) stirs.Stirred this solution 60 minutes.Collected organic layer through brine wash, is transferred to round-bottomed flask with 2mL (through being calculated as the quinhydrones of 0.2mmol).This solution is placed nitrogen.In this solution, dropwise add MeSO ₃Sex change IPA (the MeSO of 0.200mmol of H ₃H is in the IPA of 0.4mL) solution.Allow the gained mixture in nitrogen, to stir 1 hour, enriched mixture at this moment, bullion is severe slurry from EtOAc.Obtain the required product of 0.112g through solid collected by filtration and drying under reduced pressure.

Embodiment 23

Formula (1) compound (0.50g, 0.8mmol, 1.0 equivalents) is dissolved among the DCM (8mL) and with 15% aqueous solution of sodium bisulfite (4mL) stirs.Stirred this solution 60 minutes.Collected organic layer through brine wash, is transferred to round-bottomed flask with 2mL (through being calculated as the quinhydrones of 0.2mmol).This solution is placed nitrogen.In this solution, dropwise add PhSO ₃Sex change IPA (the PhSO of 0.200mmol of H ₃H is in the IPA of 0.4mL) solution.Allow the gained mixture in nitrogen, to stir 1 hour, enriched mixture at this moment, bullion is severe slurry from EtOAc.Obtain the required product of 0.118g through solid collected by filtration and drying under reduced pressure.

Embodiment 24

Under the room temperature, 10% sodium sulfite anhy 96 (200mL) aqueous solution vigorous stirring 2h of the ethyl acetate solution (200mL) of 17-allyl amino-17-demethoxylation NSC 122750 (10.0g, 17.1 mmol) and prepared fresh.Color becomes glassy yellow by grape, and the reaction completion is described.Separate each layer and use the dry organic phase of sal epsom (15g).Siccative washs through ETHYLE ACETATE (50mL).In 20 minutes, the filtrating of merging is acidified to pH 2 through the ethyl acetate solution of 1.5M spirit of salt.The gained slurry is in stirring at room 1.5h.Solid separates through filtering, through ETHYLE ACETATE (50mL) washing and in 40 ℃, and 1mm Hg, dry 16h is to obtain 9.9g (91%) pale solid.Thick quinhydrones hydrogen chlorate (2.5g) is added the moisture methanolic hydrochloric acid solution of 5%0.01N (5mL) that stirs.Make gained solution clarify through filtration, use acetone (70mL) dilution then.Solid appears after 2-3 minute.Stir gained slurry 3h in envrionment temperature, stir 1h in 0-5 ℃ then.Through the filtering separation solid, through acetone (15mL) washing and dry.

Embodiment 25

Under the room temperature, 17-allyl amino-17-demethoxylation NSC 122750 (0.350g, 10% sodium sulfite anhy 96 (7mL) aqueous solution vigorous stirring 1h of ethyl acetate solution 0.598mmol) (7mL) and prepared fresh.Color becomes glassy yellow by grape, and the reaction completion is described.Separate each layer and use the dry organic phase of sal epsom (1g).Siccative washs through ETHYLE ACETATE (1mL).The organic layer that merges in stirring at room and to wherein add TRIPHOSGENE 99.5 (0.079g, 0.239mmol).Form deposition and allow the gained mixture to stir 2h.Filtering this moment solid also concentrates organic solution.Crude product obtains the required product of 17mg through the column chromatography purifying.

Embodiment 26

Under the room temperature, 17-allyl amino-17-demethoxylation NSC 122750 (0.825g, 10% sodium sulfite anhy 96 (17.5mL) aqueous solution vigorous stirring 1h of ethyl acetate solution 0.141mmol) (17.5mL) and prepared fresh.Color becomes glassy yellow by grape, and the reaction completion is described.Separate each layer and use the dry organic phase of sal epsom (1g).Siccative washs through ETHYLE ACETATE (1mL).The organic layer that merges in stirring at room and to wherein add bromoacetyl chloride (0.222g, 1.41mmol).Form deposition and allow the gained mixture to stir 12h.Filtering this moment solid also concentrates organic solution.Crude product is dissolved in 1: 1 the THF/ water mixture (16mL).Add Na ₂CO ₃(10 equivalent) and strenuous vibration gained mixture 1hr.Use saturated NaHCO ₃Quencher should be reacted, and used brine wash, through MgSO ₄Dry and the concentrated required product of 1.1mg that obtains.

Embodiment 27

NSC 122750 25

NSC 122750 (1.12g, 2mmol, 1 equivalent) is added among the anhydrous DCM (5mL).With NH ₃MeOH solution add in this solution (9mL, 100mmol, 50 equivalents) and allow to stir 24 hours.At this moment, reaction soln is diluted through DCM, through water, with after rare HCl extraction.Collected organic layer, through brine wash, Na ₂SO ₄Dry and the concentrated purple solid that obtains.This solid obtains 17-amino-17-demethoxylation NSC 122750 of 0.239 through twice recrystallization in acetone/heptane.

17-amino-17-demethoxylation NSC 122750 (0.55g, 1mmol, 1 equivalent) is dissolved among the EtOAc (100mL).Add 10% aqueous solution of sodium bisulfite (10mL) of prepared fresh and stir 1h in envrionment temperature.Color becomes glassy yellow by grape, shows the reaction completion.Separate each layer and through dried over mgso.Siccative is through ETHYLE ACETATE washing (2 * 10mL).The filtrating that merges is acidified to pH 2 through ETHYLE ACETATE (1mL) solution of 1.5M spirit of salt in 20min.The gained slurry stirs 1.5h in envrionment temperature.Solid separates through filtering, ETHYLE ACETATE (10mL) washing, and vacuum-drying obtains product (0.524g, 87% yield).

Embodiment 28

NSC 122750 (0.500g, 0.892mmol, 1 equivalent) is dissolved in THF (10mL) 3-amino-1,2-Ucar 35 (0.813g, 8.92mmol, 10 equivalents).Stir and to react 64 hours.Use HCl quencher reaction then and extract through EtOAc.Collected organic layer is through MgSO ₄Dry also concentrating under reduced pressure.Thick material obtains the amino substituted NSC 122750 of 17-of 27mg through column chromatography purification.

17-amino geldanamycin mycin (.200g, 0.323mmol, 1 equivalent) is dissolved among the EtOAc (4mL) and uses 10% Na of prepared fresh ₂S ₂O ₄The aqueous solution (4mL) is handled.This mixture of vigorous stirring 1 hour.Collected organic layer then.Water layer is through the EtOAc of 2 * 5mL extraction.Merge organic layer, through water washing, Na ₂SO ₄Dry.Use EtOAc (1.6M, 0.6mL) the solution-treated organic layer and stirring 20 minutes of HCl then.The concentrating under reduced pressure reaction soln obtains product (0.009g).

Embodiment 29

NSC 122750 (0.022g, 0.04mmol, 1.5 equivalents) and BODIPY-FL-EDA-HCl (0.010g, 0.026mmol, 1 equivalent) are added among the anhydrous DCM (2mL).Add DIPEA (30 μ L, 0.16mmol, 6 equivalents) and stirring reaction solution 72 hours in nitrogen.Use the DCM dilute reaction solution then, through water extraction, Na ₂SO ₄Dry also concentrating under reduced pressure.Bullion obtains the amino substituted benzoquinones of 17-through column chromatography purification.With this substance dissolves in EtOAc (20mL) and use 10% Na of prepared fresh ₂S ₂O ₄The aqueous solution (5mL) is handled.This mixture of vigorous stirring 1 hour.Collected organic layer then.Water layer is through the EtOAc of 2 * 5mL extraction.Merge organic layer, through water washing, Na ₂SO ₄Dry.Use EtOAc (1.6M, 0.6mL) the solution-treated organic layer and stirring 20 minutes of HCl then.The concentrating under reduced pressure reaction soln is to doing then.Bullion is severe slurry from EtOAc/MTBE.Solid washs through MTBE, and drying under reduced pressure obtains product (0.04g).

Embodiment 30

Anhydrous ethyl acetate (170mL) is added in the flask, add 17-AAG (8.41g, 1.44mmol, 1 equivalent) subsequently.Vigorous stirring gained purple mixture under nitrogen.The 10%Na that adds prepared fresh ₂S ₂O ₄(aq) (1.682g in the 170mL deionized water, 10.1mmol, 7 equivalents) and vigorous stirring mixture 70min.Color is become orange by purple, show the reaction completion.Allow to separate each layer and use separating funnel to remove the water layer of bottom.Organic layer is through MgSO ₄Dry.Siccative is through removing by filter.Filtrating is transferred to the rotary evaporation flask.Use ETHYLE ACETATE (50mL) portioning washing MgSO ₄Filter cake, the filtrating of washing also adds the rotary evaporation flask.

On Rotary Evaporators, concentrate orange-brown mixture and obtain oily matter.

When concentrating this mixture, the ethyl acetate solution of preparation 5.3M HCl.ETHYLE ACETATE added to be blown into HCl gas 1h (through the cooling of acetone/dry ice) in the Erlenmeyer flask and in stirred mixture saturated to realize.Down this solution is risen to room temperature at nitrogen head space (head space).

Oily matter is dissolved in the acetone (252mL) and is transferred to equipped addition funnel, stirrer is in the reaction flask of TM and nitrogen.The filtrating and the washings that merge are acidified to pH 2.5 in 5min.The gained slurry stirs 18min in envrionment temperature, then through the filtration separate solid and through twice of washing with acetone (84mL).The gained solid obtains product through drying under reduced pressure.

Embodiment 31

Under the envrionment temperature, vigorous stirring 17-allyl amino-17-demethoxylation NSC 122750 (1.0g, 10% sodium sulfite anhy 96 (2g is in the 20mL water) aqueous solution of ethyl acetate solution 1.71mmol) (20mL) and prepared fresh 30 minutes.Color becomes glassy yellow by grape, shows the reaction completion.Separate each layer, organic layer is through sal epsom (1g) drying.Collect reaction solvent, siccative washs through ETHYLE ACETATE (1mL).The filtrating that merges is chilled to 0 ℃, uses the hydrobromic ethyl acetate solution acidifying of 1.5M until forming deposition.The gained slurry stirred 30 minutes in envrionment temperature.This solid separates through filtering, and ETHYLE ACETATE (1mL) washs and in 40 ℃, 1mm Hg is dry, and 16h obtains 0.352g (31%) pale solid.

Embodiment 32

Preparation as the 50mM Hydrocerol A of The compounds of this invention preparation damping fluid, 50mM xitix, pH3.1,2.44mM EDTA

The embodiment of formulation preparation

In order to prepare 1L preparation damping fluid, with 9.6g Hydrocerol A (USP), 8.8g xitix (USP) and 1.0g EDTA (ethylenediamino tetraacetic acid, disodium-calcium salt, duohydrate, USP) magnetic stirring bar with tetrafluoroethylene-coating adds in the 1L volume flask.

Add the whole volumetrical 90-95% of sterile water for injection to flask.Vigorous stirring solution is with the dissolving all solids.Use NaOH solution that the pH of damping fluid is transferred to 3.1.Add WFI to final volume.This damping fluid filters the unit vacuum filtration through 0.2 micron.Before using, solution is through nitrogen wash 1-2h.The said preparation damping fluid in closed container, 4 ℃, under nitrogen, store.

The preparation for preparing medicines:

Under the nitrogen head space, in the water cold sleeve container, use the preparation damping fluid control dissolved solids compound 15 of the nitrogen wash of precooling to prepare medicines in 4 ℃.Compound 15 solution of preparation store under the nitrogen head space in 4 ℃.

Provided the embodiment of obtaining liq medicine below .

Fill solid chemical compound 15 (500mg) and use nitrogen wash to the 10mL volumetric flask.The preparation damping fluid through (50mM Hydrocerol A, 50mM xitix, 2.44mM EDTA, pH 3.1) through nitrogen wash until dissolved oxygen level＜0.5mg/L, and in cooled on ice.Add in the volumetric flask a part of damping fluid (about 5-7mL) and high vibration dissolves until all solids.The 10mL scale that adds damping fluid to volumetric flask then.Solution is placed maintenance cooling as far as possible on ice.Use has that the 10mL syringe (Millipore, Durapore membrane, 0.2 micron) of injection filter will be clarified, brown solutions is filtered to (USP Type I) in the vial.Compound 15 solution of being prepared store under the nitrogen head space for 4 ℃.

Provided the embodiment of preparation solid form medicine below .

The 52.50g sterilized water is added 100mL to be equipped with in the flask of magnetic stirring bar.In the 100mL flask, add the 6.305g citric acid monohydrate and stir the gained mixture and be dissolved in the solution until all Hydrocerol As.In this 100mL flask, add 5.284g L-xitix then and stir and be dissolved in solution until all xitix.Then the 0.600g calcium disodium edathamil is added in this 100mL flask and stirs the gained mixture and be dissolved in solution until all calcium disodium edathamils.Then through slowly adding the pH value to 3.1 of 5M aqueous sodium hydroxide solution regulator solution.This solution was through filtering (20,0.22 microns durapore of Millipak) nitrogen wash 2 hours then.Then in stirring, nitrogen down cooling 52.04g through the solution to 0 of flushing ℃.Add 2.80g compound 15, stir the gained mixture until all compounds 15 of dissolving.This solution warp 0.22 micron pore size Durapore Millipak 200 strainers are in 0 ℃ of sterile filtration.Use through filtering (20,0.22 microns durapore of Millipak) nitrogen and be full of the head space of storage receptacle.

Then storage receptacle is placed Freeze Drying Equipment, its precooling is to-40 ℃.Cryodesiccation chamber is at-40 ℃, and 1atm kept 3 hours.The pressure of refrigeration chamber is even 100 little (micron) that fade in 1 hour.Fade to-20 ℃ and keep 100 little vacuum with the temperature of this chamber is even in 2 hours then.Fade to 0 ℃ and keep 100 little vacuum with the temperature of this chamber is even in 2 hours then.Fade to 0 ℃ and keep 100 little vacuum with the temperature of this chamber is even in 2 hours then.Then with the temperature of this chamber even fading to+10 ℃ and keep 100 little vacuum in 2 hours.Then with the temperature of this chamber even fading to+20 ℃ and keep 100 little vacuum in 2 hours.100 little vacuum were kept 48 hours and kept to the temperature of this chamber in+20 ℃.Use this chamber of nitrogen wash then and clog the container that comprises preparation with stopper.Said preparation is in-20 ℃ of storages.

Embodiment 33

(the W/V γ-Huan Hujing is as the preparation of the preparation damping fluid of The compounds of this invention for 75mM Hydrocerol A, 170mM xitix, pH 3.0,2.44mM EDTA, 1%

Formulation preparation embodiment:

In order to prepare 1L preparation damping fluid, with the 14.4g Hydrocerol A, the 30g xitix, 10g γ-Huan Hujing (cyclooctaamylose) and 1.0g EDTA add in the 1L volumetric flask with the magnetic stirring bar of teflon-coated.Add the whole volumetrical 90-95% of Injectable sterile water to flask.This solution of vigorous stirring is to dissolving all solids.Use sodium hydroxide solution (NF level) to regulate this pH of buffer value to 3.0.Add WFI to whole volume.Damping fluid is through 0.2 micron filtering unit vacuum filtration.Before using, solution is through nitrogen wash 1-2h.Storage preparation damping fluid in closed container, under 4 ℃, nitrogen.

The preparation for preparing medicines:

Under the nitrogen head space, use the preparation damping fluid control dissolved solids compound 15 of the nitrogen wash of precooling to prepare medicines in 4 ℃.Compound 15 solution of preparation store under the nitrogen head space in 4 ℃.

Embodiment 34

50mM Hydrocerol A, 25mM xitix, 1% (v/v) Tween-80,0.1% (w/v) EDTA, pH 3.0 is as the preparation of the preparation damping fluid of The compounds of this invention

Formulation preparation embodiment:

In order to prepare 1L preparation damping fluid, 9.6g Hydrocerol A, 4.4g xitix, 10mL Tween-80 and 1.0g EDTA (YD 30, disodium-calcium salt, duohydrate) are added in the 1L volumetric flask with the magnetic stirring bar of teflon-coated.Add the whole volumetrical 90-95% of Injectable sterile water (WFI) to flask.This solution of vigorous stirring is to dissolving all solids.Use sodium hydroxide solution to regulate this pH of buffer value to 3.0.Add WFI to whole volume.Damping fluid is through 0.2 micron filtering unit vacuum filtration.Before using, solution is through nitrogen wash 1-2h.In closed container, 4 ℃, nitrogen is storage preparation damping fluid down.

The preparation for preparing medicines:

Use the preparation damping fluid control dissolved solids compound 15 of nitrogen wash to prepare medicines.Compound 15 solution of preparation store under the nitrogen head space in 4 ℃.

Embodiment 35

The material of analyzed in vitro and method

Cell cultures

Human carcinoma cell line SKBR ₃, MV4-11, K562, SK-MEL-28, LnCAP and MDA-MB-468 derive from the American Type Culture Collection (Manassas, VA).Multiple melanoma RPMI-8226 and MM1.s cell from Dr.Teru Hideshima (Jerome Lipper Multiple Myeloma Center, Dana Farber CancerInstitute, Boston, MA, USA.).All cells system all is confirmed as and does not contain mycoplasma.Cell is maintained at and is supplemented with 10% hot deactivation FBS, in the RPMI-1640 substratum of 50 units/mL Streptomycin sulphate and 50 units/mL penicillium mould, at 5%CO ₂In hatch in 37 ℃.Before bed board is used for test, use to contain 0.05% trypsinase and 0.02%EDTA and phosphate buffered saline buffer (PBS) the separation of synechia cell of calcic and magnesium not.

Analyzed in vitro

The MM1.s cytotoxicity

Alamar is blue to be measured.Use the test compounds that increases concentration to hatch MM1.s cell (50,000/ hole) 72h.It is blue and measure fluorescence in 37 ℃ after hatching 4h in each hole, to add Alamar.

The SKBR3 cytotoxicity

Use the test compounds that increases concentration to hatch SKBR ₃Cell 72h.In order to carry out viability research, after adding Alamar indigo plant and hatching 6h each hole is counted.

The MDA-MB-468 cytotoxicity

Use the test compounds that increases concentration to hatch MDA-MB-468 cell 72h.In order to carry out viability research, after adding Alamar indigo plant and hatching 6h each hole is counted.

The MV4-11 cytotoxicity

Use the test compounds that increases concentration to hatch the MV4-11 cell 3 days.Use the blue counting of Alamar to estimate cell survival.

The K562 cytotoxicity

Use the test compounds that increases concentration to hatch the K562 cell.Use the blue counting of Alamar to estimate cell survival.

The SK-MEL-28 cytotoxicity

In the SK-MEL-28 of substratum cell, add to increase the

test compounds

2,3 or 4 days of concentration, use the blue cell survival of estimating of Alamar.

The LnCAP cytotoxicity

In the LnCAP of substratum cell, add to increase the test compounds 4 days of concentration, use the blue cell survival of estimating of Alamar.

Embodiment 36

17-AAG and compound 15 combine to measure for the competition of HSP90

Material

Separation from the natural human Hsp90 albumen of HeLa cell (SPP-770), reorganization dog Grp94 (SPP-766) and recombinant human Hsp70 (ESP-555) available from StressgenBiotechnologies (Victoria, BC).Complete proteinase inhibitor table derive from RocheDiagnostics (Indianapolis, IN).All other compounds and reagent are available from Sigma-Aldrich and be AG or more senior.

FP combines to measure--and BODIPY-GDM is incorporated into purifying protein

Based on Llauger-Bufi etal. (Llauger-Bufi L, Felts SJ, Huezo H; RosenN; Chiosis G.Synthesis of novel fluorescent probes for the molecularchaperone Hsp90.Bioorg Med Chem Lett (2003) 13:3975-3978) and Kim et al. (Kim J, Felts S, Llauger L; He H; Huezo H, Rosen N, Chiosis G.Development of a fluorescence polarization assay for themolecular chaperone Hsp90.J Biomol Screening (2004) 9:375-381) revised method.Combine to measure damping fluid [20mM HEPES-KOH, pH7.3,1.0mM EDTA, 100mM Repone K, 5.0mM magnesium chloride, 0.01%NP-40,0.1mg/mL bovine (BGG), 1.0mM DTT and Complete at FP from the DMSO storing solution of 20 μ M ^TMProteinase inhibitor] middle prepared fresh 20nM BODIPY-GDM solution.This solution of 10 microlitres is scattered in black round bottom 384-hole microplate (Corning#; 3676) in each hole.Add equal-volume then and combine to measure in the damping fluid one by one at FP that the human Hsp90 solution of dilution obtains final concentration 10nM BODIPY-GDM and the Hsp90 of concentration in 6.25 μ M-0.10 μ M variation.The concentration of final DMSO is 0.05%.In 30 ℃ hatch 3h after, equipped 485nm exciter filter and 535nm P/S the emission spectral filter (Perkin Elmer, Boston measure fluorescence anisotropy on EnVision 2100 multiple labeling plate telltales MA).

The competition of 17-AAG and analogue

At first with 17-AAG and compound 15 be dissolved among the DMSO with form 5.0 with the storing solution of 1.0mM concentration.Prepared fresh is from each series of compounds diluent of 20 μ M-0.20 μ M in FP combination mensuration damping fluid.Also combine to measure and prepare the solution (0.10%DMSO) that comprises 20nMBODIPY-GDM and 80nM Hsp90 in the damping fluid at FP.In the microplate of 384-hole, the solution that 10 μ L comprise BODIPY-GDM and Hsp90 mixes with equal-volume series of compounds diluent to form final concentration 10nM BODIPY-GDM, the particular compound of 40nM Hsp90 and 10 μ M-0.10 μ M different concns.Maximum DMSO concentration is 0.25% in final mensuration mixture.In 30 ℃ hatch 3h after, on EnVision 2100 multiple labeling plate telltales, measure fluorescence anisotropy.

The LabMaster glove box (M.Braun, Stratham, NH) in, measure under the nitrogen.Typically, the FP of 50mL combines the mensuration damping fluid to vacuumize through recirculation and the deoxidation with the hydrogen flushing.Refrigerating fulid input glove box with the compound storing solution among protein solution and the DMSO.In glove box, carry out all dilutions and mixed determining component subsequently as stated.In 30 ℃ hatch 3h after, from glove box, take out microplate, and on EnVision2100 multiple labeling plate telltale, measure fluorescence anisotropy immediately.

Data analysis

BODIPY-GDM and Hsp90 combine to cause increasing simultaneously fluorescence anisotropy (FA) and intensity (FI).For calculating K d, with the binding curve of the logistic function of four parameters coupling FI with respect to Hsp90 (monomer) concentration.

FI = {FI}_{\min} + \frac{({FI}_{\max} - {FI}_{\min})}{1 + {({EC}_{50} / {[E]}_{total})}^{Hill}}

For easy, regulation Hill parameter is 1.According to the value of FImax (binding partner) and FImin (free ligand), calculate the Q factor as follows:

Q = \frac{{FI}_{\max}}{{FI}_{\min}}

Use SCIENTIST program and the binding curve of following equality coupling FA subsequently with respect to Hsp90 concentration:

K_{d} = \frac{{[E]}_{free} \times {[L]}_{free}}{[EL]}

= \frac{({[E]}_{total} - [EL]) \times ({[L]}_{total} - [EL])}{[EL]}

The weight that FA is expressed as free and the contribution of combining form part with:

FA = \frac{{FA}_{\min} \times {[L]}_{free} + {FA}_{\max} \times Q \times [EL]}{{[L]}_{free} + Q \times [EL]}

= \frac{{FA}_{\min} \times ({[L]}_{total} - [EL]) + {FA}_{\max} \times Q \times [EL]}{({[L]}_{total} - [EL]) + Q \times [EL]}

Analyze the competition binding curve in a similar manner.Reduction as the FI of the function that increases inhibitor concentration is described in logistic function:

FI = {FI}_{\min} + \frac{({FI}_{\max} - {FI}_{\min})}{1 + {({{[I]}_{total} / EC}_{50})}^{Hill}}

The implicit function coupling FA that uses the competition binding equilibrium subsequently with respect to the curve of inhibitor concentration to provide Ki:

{[E]}_{total} = {[E]}_{free} + [EL] + [EI]

= {[E]}_{free} + \frac{{[E]}_{free} \times {[L]}_{total}}{{[E]}_{free} + K_{d}} + \frac{{[E]}_{free} \times {[I]}_{total}}{{[E]}_{free} + K_{i}}

[E] total, [L] total and Kd given value have been provided.

General introduction

This evidence quinone and quinhydrones Ansamycin (like 17-AAG and compound 15) all are active HSP90 suppressor factor.

Embodiment 37

The body inner analysis

Many melanoma model

In many melanoma cell series of the intravital mankind of male SCID/NOD mouse RPMI-8226, studied the effect of test compounds.In this research, to male mice subcutaneous transplantation RPMI-8226 cell (1 * 10 ⁷Individual cell).When the average tumor size reaches 100mm ³The time, animal is divided into treatment group (N=10-15/ group) at random thus accepting carrier (50mM Hydrocerol A, 50mM xitix, 2.4mM EDTA, regulate pH value to 3.0) or 100mg/kg (300mg/m weekly in continuous three days ²) test compounds.(IV) gives tester or carrier in the tail cava vein, gives 0.2mL in about 20 seconds.Put to death animal and comparison of tumor volume after 45 days.

Breast cancer model

In the MDA-MB-468 breast cancer model, study the ability that reduces the Subcutaneous tumor load with the evaluation test compound.In this research, female nu/nu athymic mouse is by subcutaneous transplantation MDA-MB-468 cell (1 * 10 ⁷Individual cell).When the average tumor size reaches 100mm ³The time, animal is divided into (N=10-15/ group) following treatment group at random: semiweekly carrier or 100mg/kg (300mg/m ²) test compounds.(IV) gives tester or carrier in the tail cava vein, gives 0.2mL in about 20 seconds.Put to death animal and comparison of tumor volume after 120 days.

The ovarian cancer model

In SKOV-3 ovary mouse heteroplastic transplantation model, study the ability that reduces the Subcutaneous tumor load with the evaluation test compound.In this research, female nu/nu athymic mouse is by subcutaneous transplantation SKOV-3 cell (1 * 10 ⁷Individual cell).When the average tumor size reaches 100mm ³The time, animal is divided into (N=10-15/ group) treatment group at random to accept carrier or semiweekly 100mg/kg (300mg/m ²) test compounds.(IV) gives tester or carrier in the tail cava vein, gives 0.1mL in about 10 seconds.Put to death animal and comparison of tumor volume after 88 days.

Mouse lewis lung carcinoma model

In mouse lewis lung carcinoma model, study the ability that reduces Subcutaneous tumor load and lung transfer incidence with the evaluation test compound.In this research, the C57B1/6 mouse is by subcutaneous transplantation lewis lung carcinoma cell (1 * 10 ⁶Individual cell).When the average tumor size reaches 71mm ³The time, animal is divided into (N=10-15/ group) following treatment group at random: Monday, Wednesday and Friday, (MWF) accepted carrier or 75mg/m ²Compound 15 continued for three weeks.(IV) gives tester or carrier in the tail cava vein, gives 0.2mL in about 20 seconds.Put to death animal and comparison of tumor volume after 25 days.

Prostate cancer

In mouse PC-3 prostate cancer heteroplastic transplantation model, carry out reducing when two researchs are united as single medicine or with present standard care medicine with the evaluation test compound ability of Subcutaneous tumor load.In two researchs, male nu/nu athymic mouse is all by subcutaneous transplantation PC-3 cell (1 * 10 ⁷Individual cell).When the average tumor size reaches 100mm ³The time, animal is divided into (N=10-15/ group) treatment group at random.In first research, mouse is accepted carrier, 100mg/kg (300mg/m weekly for twice ²) test compounds.(IV) gives tester or carrier in the tail cava vein, gives 0.2mL in about 20 seconds.Put to death animal and comparison of tumor volume after 64 days.

In this model, carry out the associating of second research with evaluation test compound and other standard care medicine TX.In this research, the different windings of a random arrangement 10-15 mouse receive carrier, twice 100mg/kg (300mg/m weekly ²) test compounds, weekly TX 5mg/kg (15mg/m ²) or the associating of test compounds and TX.Put to death animal and comparison of tumor volume after 64 days.

Embodiment 38

Biological results

The BA analytical results that has shown quinhydrones of the present invention below.All value representations are MV ± SEM.Data analysis is made up of the one-way analysis of variance, in the time of suitably, with after the difference between Dnnets test evaluation carrier and the treatment group.It is significant that the difference of p＜0.05 is considered to.

In vitro results

CloneCompound 15 (EC ₅₀) 17-AAG (EC ₅₀)

MM1.s 307nM 306nM

SKBr3 32nM 34nM

MDA-MB-468 335nM 356nM

MV4-11 25nM 38nM

K562 29nM 50nM

SK-MEL-28 200nM ?------

LnCAP 73nM ------

Result in the body

Clone The correlated tumor propagation of % and carrier

Compound 15 Compound 15+ TX

RPMI-8226 71％ -------

MDA-MB-468 76％ -------

SKOV-3 59％ -------

Lewis pneumonocyte 60%-------

PC-3 50％ 84％

Compound 15 and the combination of 17-AAG to HSP90

CompoundK _j

Compound 15 28nM

17-AAG 67nM

Equivalent variations and reference are introduced

Be interpreted as embodiment as herein described and embodiment and only be illustrative purposes for example, and will point out those skilled in the art many variations or change about it, these variations or change are included in the spirit and scope of the application and accompanying claims.All publications, patent and patented claim that this paper quotes are incorporated herein by reference at this with various purposes in full.

Claims

1. suc as formula the compound shown in 3:

X wherein ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, Cyclamic Acid root, thiocyanate ion, naphthalene-2-sulfonic acid root and oxalate.

2. the compound of claim 1, the pKa of wherein pharmaceutically acceptable acid in water is-10 to 1.

3. the compound of claim 1, the pKa of wherein pharmaceutically acceptable acid in water is-10 to-3.

4. suc as formula the compound shown in 15,

5. pharmaceutical composition comprises each described compound and at least a pharmaceutically acceptable vehicle among the claim 1-4.

6. according to the pharmaceutical composition of claim 5, wherein said composition further comprises inhibitor.

7. according to the pharmaceutical composition of claim 6; Wherein said inhibitor is selected from ascorbate salt, halfcystine hydrogen chlorate, sodium sulfite anhy 96, sodium metabisulfite, S-WAT, thioglycerin, Thioglycolic acid sodium salt, sodium formaldehyde sulphoxylate, anti-bad blood acyl cetylate, butylated hydroxyanisole, DBPC 2,6 ditertiary butyl p cresol, Yelkin TTS, Tenox PG or alpha-tocopherol.

8. according to the pharmaceutical composition of claim 6, wherein said inhibitor is an ascorbate salt.

9. according to the pharmaceutical composition of claim 6, wherein said inhibitor is the L-xitix.

10. according to the compsn of claim 5, wherein said composition further comprises buffer reagent.

11. according to the pharmaceutical composition of claim 10, wherein said buffer reagent is selected from Citrate trianion, ascorbate salt, phosphoric acid salt, hydrocarbonate, carbonate, fumarate, acetate, tartrate, malate, SUMATRIPTAN SUCCINATE, lactic acid salt, maleic acid salt, glycocoll and other naturally occurring α or beta-amino acids.

12. according to the pharmaceutical composition of claim 10, wherein said buffer reagent is a Citrate trianion.

13. according to the pharmaceutical composition of claim 10, wherein said buffer reagent is a citric acid monohydrate.

14. according to the pharmaceutical composition of claim 5, wherein said composition further comprises metal chelator.

15. according to the pharmaceutical composition of claim 14, wherein said metal chelator is selected from YD 30 (EDTA), diethylene base-triamine-five-acetate (DTPA), ethylene glycol tetraacetic (EGTA), nitrilotriacetic acid (NTA), sorbyl alcohol, tartrate, N-hydroxyiminodiacetic acid salt, hydroxyethyl-YD 30,1-and 3-trimethylenedinitrilo-tertraacetic acid, 1-and 3-diamino--2-hydroxy propane four-acetate, gluconic acid sodium salt, hydroxyl ethane di 2 ethylhexyl phosphonic acid, phosphoric acid and salt thereof.

16. according to the pharmaceutical composition of claim 14, wherein said metal chelator is YD 30 (EDTA).

17. according to the pharmaceutical composition of claim 14, wherein said metal chelator is YD 30 (EDTA) calcium disodium monocalcium salt compound.

18. according to the pharmaceutical composition of claim 5, wherein said composition further comprises inhibitor, and buffer reagent.

19. according to the pharmaceutical composition of claim 5, wherein said composition further comprises inhibitor, and metal chelator.

20. according to the pharmaceutical composition of claim 5, wherein said composition further comprises buffer reagent, and metal chelator.

21. according to the pharmaceutical composition of claim 5, wherein said composition further comprises inhibitor, buffer reagent and metal chelator.

22. according to the pharmaceutical composition of claim 21, wherein inhibitor is an ascorbate salt, buffer reagent is that Citrate trianion and metal chelator are YD 30 (EDTA).

23. according to the pharmaceutical composition of claim 21, wherein inhibitor is the L-xitix, buffer reagent is that citric acid monohydrate and metal chelator are YD 30 (EDTA) calcium disodium monocalcium salt compounds.

24. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.001-0.1.

25. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.01-0.05.

26. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.001-1.

27. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.01-1.

28. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of citric acid monohydrate and formula 3 or formula 15 compounds is 0.05-2.

29. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of citric acid monohydrate and formula 3 or formula 15 compounds is 0.2-1.

30. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.001-0.1; And the molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.001-1.

31. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.01-0.05; And the molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.01-1.

32. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.001-0.1; The molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.001-1; And the molar ratio range of citric acid monohydrate and formula 3 or formula 15 compounds is 0.05-2.

33. according to the pharmaceutical composition of claim 23, wherein the molar ratio range of YD 30 (EDTA) calcium disodium monocalcium salt compound and formula 3 or formula 15 compounds is 0.01-0.05; And the molar ratio range of L-xitix and formula 3 or formula 15 compounds is 0.01-1; And the molar ratio range of citric acid monohydrate and formula 3 or formula 15 compounds is 0.2-1.

34., further contain solubilizing agent according to each pharmaceutical composition of claim 5-33.

35. according to the pharmaceutical composition of claim 34, wherein said solubilizing agent is selected from polyoxyethylene glycol sorbitan aliphatic ester, polyethylene glycol stearate, benzylalcohol, ethanol, polyoxyethylene glycol, Ucar 35, glycerine, Schardinger dextrins or Prist.

36. according to each pharmaceutical composition of claim 5-35, wherein having said formula 3 or formula 15 compound concentrations is 0.00016M-0.160M.

37. according to each pharmaceutical composition of claim 5-35, wherein having said formula 3 or formula 15 compound concentrations is 0.00032M-0.080M.

38. each each pharmaceutical composition of compound or claim 5-37 of claim 1-4 is treated the purposes in the medicine of cancer in being prepared in Mammals.

39. according to the purposes of claim 38, wherein said cancer is the cancer of hemopoietic system, immunity system, endocrine system, pulmonary system, gastro-intestinal system, Musculoskeletal, reproductive system, cns or urinary system.

40. according to the purposes of claim 38, wherein said cancer is arranged in mammiferous myeloid tissue, Lymphoid tissue, pancreatic tissue, parathyroid tissue, lung, colon, rectal tissue, anus tissue, hepatic tissue, skin, bone, ovary tissue, uterine cancer cell, cervical tissue, breast, prostate gland, testis tissue, brain, brain stem, meninges tissue, kidney or bladder.

41. according to the purposes of claim 38, wherein said cancer is arranged in mammiferous myeloid tissue, Lymphoid tissue, breast, lung, ovary or prostate gland.

42. according to the purposes of claim 38, wherein said cancer is selected from mammary cancer, multiple myeloma, carcinoma of prostate, Hodgkin lymphoma, non-Hodgkin lymphoma, acute lymphoblastic leukemia, lymphocytic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, renal cell carcinoma, malignant melanoma, carcinoma of the pancreas, lung cancer, colorectal carcinoma, colorectal carcinoma, brain cancer, kidney disease, head and neck cancer, bladder cancer, thyroid carcinoma, ovary cancer, Cervical cancer or myelodysplastic syndrome.

43. according to the purposes of claim 38, wherein said cancer is a mammary cancer.

44. according to the purposes of claim 38, wherein said cancer is an acute myelogenous leukemia.

45. according to the purposes of claim 38, wherein said cancer is a chronic lymphocytic leukemia.

46. according to the purposes of claim 38, wherein said cancer right and wrong-small cell lung cancer.

47. according to the purposes of claim 38, wherein said cancer is a small cell lung cancer.

48. according to the purposes of claim 38, wherein said cancer is an ovarian cancer.

49. according to the purposes of claim 38, wherein said Mammals is selected from primate, horse, dog, cat and ox.

50. according to the purposes of claim 38, wherein said Mammals is human.

51. according to the purposes of claim 38, the administering mode of wherein said compound or pharmaceutical composition is selected from that suction, oral, intravenously, hypogloeeis, eyes, transdermal, rectum, vagina, part, intramuscular, intra-arterial, sheath are interior, subcutaneous, oral cavity or nose administration.

52. according to the purposes of claim 38, wherein administering mode is an intravenously.

53. the preparation method of formula 3 compounds,

X wherein ^-Be selected from chlorine, bromine, iodine, H ₂PO ₄ ^-, HSO ₄ ^-, methanesulfonate, Phenylsulfonic acid root, tosic acid root, trifluoromethanesulfonic acid root, 10-camphorsulfonic acid root, naphthalene-1-sulfonic acid-5-sulfonate radical, ethane-1-sulfonic acid-2-sulfonate radical, Cyclamic Acid root, thiocyanate ion, naphthalene-2-sulfonic acid root and oxalate,

This method comprises makes formula 1 compound

In reaction solvent, mix, obtain formula 8 compounds with reductive agent:

And formula 8 compounds are mixed with pharmaceutically acceptable acid, obtain formula 3 compounds.

54. according to the method for claim 53, wherein said reductive agent is selected from sodium sulfite anhy 96, zinc, xitix or electrochemical reduction.

55. according to the method for claim 53, wherein said reductive agent is a sodium sulfite anhy 96.

56. method according to claim 53; Wherein said reaction solvent is selected from methylene dichloride, chloroform, ethylene dichloride, chlorobenzene, THF, 2-MeTHF, ether, diglyme, 1,2-glycol dimethyl ether, MTBE, THP, dioxane, 2-ethoxy butane, methyl butyl ether, ETHYLE ACETATE, methyl acetate, 2-butanone, water and composition thereof.

57. according to the method for claim 53, wherein said reaction solvent is the mixture of ETHYLE ACETATE and water.

58. according to the method for claim 53, the pKa of wherein said acid in water is-10-1.

59. according to the method for claim 53, the pKa of wherein said acid in water is-10--3.

60. according to the method for claim 53, wherein said acid is selected from HCl, HBr, H ₂SO ₄, methylsulfonic acid, Phenylsulfonic acid, tosic acid, trifluoromethanesulfonic acid, camphorsulfonic acid, naphthalene-1,5-disulfonic acid, ethane-1,2-disulfonic acid, Cyclamic Acid, thiocyanic acid, naphthalene-2-sulfonic acid or oxalic acid.

61. according to the method for claim 53, wherein said acid is HCl; And X ^-Be Cl ^-

62. according to the method for claim 53, wherein said acid is HBr; And X ^-Be Br ^-

63. according to the method for claim 53, wherein said acid adds as gas.

64. according to the method for claim 53, wherein said acid is dissolved in the organic solvent.

65. according to the method for claim 64, wherein said organic solvent is selected from ETHYLE ACETATE, methylene dichloride, Ucar 35 and dioxane.