TW201632182A - Triptolide and derivatives thereof in the treatment of tumors and precancerous pathologies of the skin - Google Patents

Abstract

The present invention concerns a compound selected from triptolide, derivatives thereof, and pharmaceutically acceptable salts of triptolide and derivatives thereof, or a composition comprising such a compound for use in the topical treatment and/or prevention of a tumor or precancerous pathology of the skin.

Description

Triptolide and its derivatives for treating tumors and precancerous pathology of the skin Field of invention

The present invention relates to the use of triptolide, a derivative thereof, and the like, and a pharmaceutically acceptable salt thereof for use in the local treatment and prevention of tumors and precancerous pathologies of the skin.

Background of the invention

Skin tumors and precancerous pathology pose serious public health problems. Some are caused by frequent exposure to ultraviolet light and can develop into a lethal form of cancer. There are essentially two types of skin tumors:

1) Cancer developed from epidermal cells of the basal layer (basal cell carcinoma) or the upper layer (squamous cell carcinoma). It is the most common, accounting for 90% of skin cancer, 75% of which are basal cells and 20% are squamous cells. Basal cell carcinoma cannot metastasize; squamous cell carcinoma metastasizes but rarely metastasizes, mainly in lymph nodes near the tumor. Cancer generally develops in several stages. With regard to squamous cell carcinoma, tumors usually begin to be damaged within the epidermis. Crust (sun keratosis) or an eczema (Bowen's disease) is formed on the surface. It invades the deeper dermis, which is a special stage of invasive cancer. Sign.

2) Melanoma develops from melanocytes, melanocytes are cells that produce melanin, and melanin is responsible for brown or red pigmentation of the skin. More specifically, there are two main types of pigments: brown, which produces a tan color and provides some UV protection, as well as unprotected red (freckle skin). Individuals that primarily produce red pigments do not tan and have a higher risk of skin cancer. Melanocytes are usually found in the epidermis and bind to epidermal cells deep in the epidermis. A "beauty spot" or sputum, a benign lesion that corresponds to the accumulation of melanocytes in the dermis, indicating why it is brown or red. Melanoma is much less common, but it may develop in young people. They must be detected and treated quickly as they spread throughout the body and produce a very difficult to treat metastasis.

Solar keratosis (AK), also known as actinic keratosis, is a skin pathology. It is the hypertrophy of the stratum corneum of the epidermis. AK is the first stage, leading to the development of squamous cell carcinoma, and thus is considered a "precancerous" lesion. Although most of the AK is benign, some studies report that approximately 10% may be converted to squamous cell carcinoma. 40 to 60% of squamous cell carcinomas result from untreated AK. Approximately 2 to 10% of squamous cell carcinomas spread to internal organs and mortality that threatens prognosis.

AK occurs in areas of the body that are often exposed to sunlight, especially to UVB rays.

It mainly affects Caucasians. For example, the prevalence of solar keratosis in Australia is estimated to be approximately 40%. In the United States, it is estimated that there are more than Eighty-two million patients have AK. The annual cost of treating AK in the United States in 2002 is estimated to be approximately $1 billion.

There are various treatment options for the treatment of AK: cryotherapy, surgery, local photodynamic therapy (PDT), laser, curettage, and electrocoagulation, topical administration containing imiquimod, 5-fluorouracil ( 5-fluorouracil), ingenol mebutate or diclofenac. The treatment should be tailored to the location and characteristics of the AK.

However, these various treatments sometimes fail to succeed, and some leave a permanent, unbeautiful flaw. There is therefore still a need for a limited, optional, effective, and rapid AK treatment of side effects.

Triptolide (TRP) and its derivatives, in particular the biguanide type, of the following formula (I) are molecules which can be extracted from traditional Chinese plants of the family Celastraceae , especially the Tripterygium wilfordii .

The present inventors have surprisingly demonstrated the efficacy of triptolide in the prevention and treatment of tumors or precancerous pathologies of the skin, especially solar keratosis. The inventors further exhibited a low level of percutaneous access to Tripterygium wilfordii The lactone is effective locally.

Summary of invention

The invention thus relates to a use of a pharmaceutically acceptable salt selected from the group consisting of triptolide, a derivative thereof, triptolide and a derivative thereof, and a mixture thereof, for topical treatment and/or Prevent tumors or precancerous pathology of the skin.

The invention further relates to the use of a topical composition comprising at least one compound of the invention and at least one pharmaceutically acceptable excipient for the treatment and/or prevention of a tumor or precancerous pathology of the skin.

The invention further relates to a method of treating and/or preventing a tumor or precancerous pathology of the skin by topically administering an effective amount of a compound or composition according to the invention to a patient in need thereof.

The invention further relates to the use of a compound or composition according to the invention for the manufacture of a topical medicament for the treatment and/or prevention of tumors or precancerous pathologies of the skin.

In the context of the present invention, "pharmaceutically acceptable" means that it can be used to prepare a pharmaceutical composition which is generally safe, non-toxic, non-biologically and otherwise undesirable, and which is acceptable for veterinary and human pharmaceutical use. .

A "pharmaceutically acceptable salt" of a compound means a salt which is pharmaceutically acceptable as defined herein and which possesses the desired pharmacological activity of the parent compound.

The pharmaceutically acceptable salts specifically include: (1) with a pharmaceutically acceptable inorganic acid such as hydrochloric acid, hydrogen a pharmaceutically acceptable acid addition salt formed by bromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with a pharmaceutically acceptable organic acid such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, Citric acid, ethanesulfonic acid, fumaric acid, grape heptanoic acid, gluconic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid , muconic acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, benzoyl-L-tartaric acid, tartaric acid, p-toluenesulfonic acid, trimethylacetic acid, trifluoroacetic acid and the like a pharmaceutically acceptable acid addition salt formed, and (2) when the protonic acid present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion; or Accepted organic base coordination, such as diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, and the like; or with a pharmaceutically acceptable inorganic base Coordination, for example, aluminum hydroxide, calcium hydroxide, hydrogen and oxygen A pharmaceutically acceptable base addition salt formed by the formation of potassium, sodium carbonate, sodium hydroxide and the like.

When the compound contains the function of an acid, the compound may be a sodium salt.

In accordance with the present invention, "triptolide derivative" means all of the triptolide derivatives having equivalent or greater potency than triptolide. The efficacy of triptolide and its derivatives can be measured, inter alia, by assessing the efficacy of, for example, the variation in the number of solar keratosis lesions during the performance test, especially in the examples below. According to "efficacy equivalent to" means triptolide derivative It will have an efficacy between 50 and 100% of the efficacy of triptolide, preferably between 80 and 100%. Thus, "efficacy equivalent to or greater than" means that the triptolide derivative will have a potency greater than or equal to 50% of the efficacy of triptolide, preferably. 80% performance.

The triptolide derivative according to the present invention is as described in the article by Ma et al. (2007) Phytochemistry 68: 1172-1178.

In one embodiment, the triptolide derivative is selected from the group consisting of or the following: triptonide, tripdiolide, triptolide Triptolidenol, 16-hydroxytriptolide, triptriolide, 12-epitriptriolide, 14-epitriprin (14-epitriptolide), tripoxalide and derivative PG-490-88, especially in Liu, Q et al. (2004) Biochem. Biophys. Res. Commun. 319: 980-986. The sodium salt form of the formula:

In another embodiment, the triptolide derivative according to the present invention is capable of activating terpenoid metabolites, such as those cited in particular by Brinker et al. (2007) Phytochemistry 68: 732-766, including Leigong Wilfordine, euponine, and cangorinine.

In particular, the triptolide derivative according to the present invention may be selected from a triptolide type triptolide derivative selected from the group consisting of or consisting of triptolide carbonyl lactone ( Triptonide), tripdiolide, triptolidenol, 16-hydroxytriptolide, triptriolide, 12-table tripterygium 12-epitriptriolide, 14-epitriptolide and derivative PG-490-88. In particular, the triptolide derivative according to the present invention may be selected from the group consisting of triptonide, tripdiolide and the derivative PG-490-88.

In another embodiment, the compound according to the present invention is triptolide, triptonide, tripdiolide, derivative PG-490-88 or pharmaceutically acceptable thereof. Salt.

In another embodiment, the compound according to the invention is triptolide, derivative PG-490-88 or a pharmaceutically acceptable salt thereof, especially triptolide or derivative PG-490-88 Sodium salt.

In another embodiment, the compound according to the present invention is triptolide, triptonide, tripdiolide or a pharmaceutically acceptable salt thereof, particularly in tripterygium wilfordii Ester, tripterygium carbonyl lactone or triptolide. Triptolide, Tripterygium wilfordii Lactone and triptolide are very similar in structure and have comparable activities.

In another embodiment, the compound according to the invention is triptolide or a pharmaceutically acceptable salt thereof, especially triptolide.

The triptolide, its derivatives, and the pharmaceutically acceptable salts thereof, such as the present invention, can be obtained by all methods known to those skilled in the art.

The composition according to the present invention may comprise triptolide and/or one or more triptolide derivatives and/or pharmaceutically acceptable salts thereof in purified form or in the form of extracts, such as, for example, Can be crude or concentrated.

By "purified form" in the context of the present invention is meant a compound comprising at least 90% by weight, in particular at least 95% by weight, in particular at least 99% by weight, of triptolide, triptolide derivative or It is a pharmaceutically acceptable salt thereof.

For example, purified forms of triptolide, derivatives thereof, and the like, which are pharmaceutically acceptable salts, can be obtained by chemical synthesis (Miller NA et al, Chem Commun (Camb). 2008 Mar 14; 10): 1226-8). Optional, the purified forms of the triptolide, derivatives thereof, and the pharmaceutically acceptable salts thereof from the Celastraceae (family Celastraceae) plants, especially Tripterygium (Tripterygium) plants, such as triptolide ( Tripterygium wilfordii ) or Tripterygium regelii , extracted and purified.

Purification can be carried out by liquid chromatography. One method for obtaining purification to more than 95% of triptolide is described, inter alia, in Ma et al. (2007) Phytochemistry 68: 1172-1178.

In another embodiment, the triptolide, a derivative thereof, and the like and a pharmaceutically acceptable salt thereof according to the present invention may be in the form of an extract after being extracted from a family of the family Celastraceae . In particular, Tripterygium plants, such as Tripterygium wilfordii or Tripterygium regelii . The extract may be crude or concentrated in terms of triptolide, triptolide derivatives and/or pharmaceutically acceptable salts thereof. The extract may optionally comprise components (including active ingredients) from the plant in addition to the pharmaceutically acceptable salts of triptolide, derivatives thereof and the like. A "crude extract" is an extract obtained directly from a plant. By "concentrated extract" is meant that, in particular, in relation to the crude extract, the amount of triptolide, its derivatives and/or its pharmaceutically acceptable salts exceeds 30% by weight. In particular, more than 50% by weight of an extract. In particular, the triptolide-concentrated extract according to the invention can be obtained by the method described in international application WO2011/054929.

Purified, crude or concentrated extracts according to the invention may be obtained from any part of the family of the family Celastraceae , especially the roots. Optionally, they can be obtained from plant cell cultures of such plants. In the case of plant cell cultures, such extracts can be obtained, in particular, from suspensions of such cell cultures (for example as described in WO 2011/054929).

In one embodiment, the compound according to the present invention, that is, triptolide, triptolide derivative or a pharmaceutically acceptable salt thereof, is in a purified form.

The term "tumor or precancerous pathology of the skin" particularly means a tumor or precancerous pathology selected from the group consisting of the following or a group consisting of: ultraviolet-induced tumor or precancerous pathology, metastasis, And T-cell lymphoma of the skin.

In one embodiment, the ultraviolet-induced tumor or precancerous pathology of the skin is particularly selected from the group consisting of or consisting of: solar keratosis, basal cell carcinoma, melanoma, and squamous cell carcinoma .

In another embodiment, the pathology of pre-cancerous skin induced by ultraviolet light according to the present invention is solar keratosis.

All of these skin tumors and precancerous pathologies are known to those skilled in the art, and those skilled in the art will readily understand how to recognize and distinguish one another.

In particular, solar keratosis can be characterized by the appearance of solar keratosis lesions. According to the "solar keratosis injury" line means the hypertrophy of the upper layer of the epidermis, resulting in thickening of the stratum corneum. The first period is characterized by a variable size (in a centimeter scale) and red spots with rough surfaces and inaccurate boundaries. At a later stage, the thickening of the stratum corneum becomes more pronounced (yellow or brown hyperkeratosis, If you try to remove it, it will bleed). Diagnosis is usually clinical, but histological examination (skin section) provides a diagnosis (overly atrophic or hyperacanthotic epidermis composed of numerous dysplastic dysplastic keratinocytes, hyperkeratosis and parakeratosis) The keratinization appears).

The squamous cell carcinoma according to the present invention preferably derives from the existing solar keratosis lesions.

Microscopic lesions that can only be seen through histology appear before the visible damage to the naked eye (visible to the naked eye).

For the purposes of the present invention, "metastasis" means the transfer of localized skin of any type of tumor, such as a breast or lung tumor, or a metastasis of melanoma.

In the context of the present invention, "treatment" means "inhibition of the development of a tumor or precancerous pathology of the skin according to the invention, more particularly degradation, preferably disappearance.

For the purposes of the present invention, "preventing" means preventing or delaying the onset of a tumor or precancerous pathology of the skin according to the invention.

The treatment or prevention according to the invention is suitable for use in humans or animals.

When the precancerous pathology according to the present invention is solar keratosis, the term "treatment of solar keratosis" means, in particular, inhibition of the occurrence of new solar keratosis damage and suppression of existing sunlight. Development of keratosis lesions (especially from microscopic development to macroscopic), inhibition of the growth of existing macroscopic keratosis lesions, degradation of existing solar keratosis lesions, and/or The disappearance of existing solar keratosis damage.

In the context of the present invention, "partial" means The composition or composition can be administered by application to the surface of the skin or to the mucosa.

The topical compositions according to the invention may, in particular, be in any form which permits topical application: creams, gels, ointments, patches and the like. Preferably, the composition according to the invention will be in the form of a cream.

In one embodiment, the topical composition according to the present invention comprises triptolide and/or one or more of its derivatives, at a concentration of 0.0002% by weight and 2% by weight relative to the weight of the final composition. Between 0.001% by weight and 2% by weight, or between 0.001% by weight and 1% by weight, or between 0.01% by weight and 1% by weight, or between 0.01% by weight Between % and 0.1% by weight, or between 0.005 and 0.1% by weight. Triptolide and/or one or more of its derivatives may be in the form of a pharmaceutically acceptable salt.

In another embodiment, the topical composition according to the present invention comprises triptolide having a concentration of between 0.0002% and 2% by weight, for example between 0.001% by weight, relative to the weight of the final composition. Between 2% by weight, or between 0.001% and 1% by weight, or between 0.01% and 1% by weight, or between 0.01% and 0.1% by weight, or It is between 0.005 wt% and 0.1 wt%. Triptolide can be in the form of a pharmaceutically acceptable salt.

In another embodiment, the dose of triptolide and/or one or more of its derivatives is administered between 1 μg and 1.5 mg/cm ² of skin, or between 5 μg and Between 1.5 mg/cm ² of skin, for example, between 1 μg and 1 mg/cm ² of skin, or between 10 μg and 1 mg/cm ² of skin. Triptolide and/or one or more of its derivatives may be in the form of a pharmaceutically acceptable salt.

The compound or composition according to the present invention may be administered once or more per day, and the duration of the treatment may vary depending on the severity of the tumor or precancerous pathology of the skin to be treated, and may be known to the practitioner or practitioner. Easy to adjust. In one mode of administration, the composition will not be administered more than 1, 2, 3, 4 or 5 times, for example more than 3 times.

In accordance with the invention, "pharmaceutically acceptable excipient" means an excipient which, when administered to an animal or human, is compatible with the other ingredients of the composition, and which does not cause side effects. , an allergic reaction or other unintended reaction.

In the context of the present invention, "excipient" means especially one or more surfactants, for example polyethylene glycol, hydroxystearate, ethoxylated fatty acid esters, B. Oxidized fatty alcohol; one or more solvents such as, for example, octyldodecanol, propylene glycol dicaprylocaprate; one or more water soluble polymers such as PVP, hyaluronic acid or hyaluronic acid Sodium; one or more thickeners such as, for example, natural or semi-synthetic gums; one or more gelling agents, for example carbomer; one or more inorganic fillers, for example zinc oxide, talc Powder, clay; one or more emulsifiers, such as cetearyl alcohol; one or more preservatives, for example phenoxyethanol; one or more antimicrobial agents; one or more preservatives (antiseptics) One or more antioxidants, for example tocopherol acetate; one or more chelating agents, for example EDTA; Or a plurality of pigments; one or more fragrances; one or more color formers; one or more pH adjusting agents, for example, salts, acids, bases; or mixtures thereof.

In one embodiment, the composition is such that the compound according to the invention does not penetrate the skin. Penetrating the skin can be measured by a test using a Franz static diffusion cell, especially as the OECD guidelines for Testing of Chemicals (New Guidelines 428, Skin Absorption: In Vitro Method) 13 April 2004)). When more than 60%, for example more than 80% or 90% of the compound according to the invention is administered, the compound according to the invention is considered to not penetrate the skin.

Figure 1: Average number of solar keratosis lesions before and after treatment with triptolide in various doses (0 μg (vehicle), 50 μg or 150 μg per mouse) and in or outside the treated area .

Figure 2: Photographs of mouse skin treated with various doses (0 μg (vehicle), 50 μg or 150 μg per mouse) of triptolide over time.

Figure 3: Daylight in the treated area before and after treatment with triptolide (10 μg or 50 μg per mouse), triptolide PCC (10 μg or 50 μg per mouse) or vehicle (control) The average number of sexual keratosis injuries.

Figure 4: Triptolide (10 μg or 50 μg per mouse), triptolide PCC (10 μg or 50 μg per mouse) or vehicle over time (Control) Photograph of the skin of the treated mice.

Detailed description of the preferred embodiment Example: Example 1: Mouse study 1. Materials and methods 1.1. Induction of solar keratosis

Female non-selfed, hairless SKH1 mice (Charles River Laboratories) between 6 and 8 weeks old, exposed to UVB rays daily for 14 weeks. The UVB intensity, confirmed by a luminosity meter, was 50 mJ/cm ² /day for 10 days, then 55 mJ/cm ² /day for 10 days, and the last remaining period was 60 mJ/cm ² /day. The dose of 50 mJ/cm ² /day was preliminarily determined to be the smallest erythema dose in the mouse strain.

The mice gradually develop microscopic and then damage to the visible keratosis of the sun. These lesions are characterized by histological and anatomopathological levels of solar keratosis lesions comparable to human solar keratosis lesions.

1.2. Products used Reference product/model verification

Two reference products for the treatment of sun keratosis damage in humans, ingenol mebutate (0.015%, Picato ^® ) and 5-fluorouracil (0.5%, Carac®), Local administration to animals (3 to 5 per group, relative to a control group receiving only vehicle), and the therapeutic activity of the mouse model demonstrating this solar keratosis, pharmacologically validated this model appropriately Sex.

Triptolide and triptolide PCC

Triptolide Research (UK) was formulated in a carrier consisting of 5% DMSO, 70% glycerol, and 25% water. Triptolide PCC (plant cell culture extract) was formulated in a carrier consisting of 5% DMSO, 50% glycerol, and 45% water. The triptolide PCC line was purified from a meristematic cell culture from Tripterygium wilfordii plants as described in application WO 2011/054929 (Examples 1 to 7). Triptolide is a composition containing triptolide (triptolide or triptolide PCC) at 0.005% (or 10 μg per animal) or 0.025% (or 50 μg per animal) Or a dose of 0.075% (or 150 μg per animal) is administered to the animal.

1.3. Locally agreed agreements

The composition to be evaluated (placebo (vehicle alone) or containing 0.005% or 0.025% or 0.075% triptolide) was administered topically to the mice to assess their presence at the beginning of treatment. The therapeutic effects of sunken keratosis lesions and their prophylactic effects on visible damage to the naked eye.

At the beginning of treatment (weeks 16-17 after the onset of UVB exposure), only mice with visible solar keratosis lesions were selected and randomized between treatment and control groups according to the number of lesions visible to the naked eye. The group consisted of between 3 and 5 mice. UVB exposure was discontinued at the beginning of treatment.

The product was placed in a closed manner in a chamber having a diameter of 19 mm (or 2.83 cm ² ), placed and taped to the defined area of each mouse. The composition containing triptolide was applied to the skin for 30 hours as a single closed application. The occlusive dressing was removed after 30 hours and the animals were closely watched for 4 weeks (17th to 20th week after the start of UVB irradiation). The triptolide-free vehicle was administered in the same manner as a placebo control.

1.4. Evaluation of therapeutic efficacy Clinical tracking

The weight of the animal was weighed three times a week between the 17th week and the 21st week. Photographs were taken once a week for the dorsal and left lateral and right lateral surfaces of each animal to pay close attention to changes in solar keratosis injury. Count the damage to solar keratosis within and outside the treated area, and pay close attention to the progression of the injury and the efficacy of the treatment.

Histological analysis

Skin from the treated and untreated areas of each animal was sectioned for histological and immunohistochemical analysis at study termination (week 21). Thus, the presence of solar keratosis lesions was confirmed by histology and compared between groups. The level of epidermal cell proliferation was assessed by Ki67 labeling.

2. Results 2.1 clinical tracking

Topical treatment was perfectly tolerated and triptolide had no effect on animal weight. This indicates that the animal is extremely well tolerated by triptolide administered via this route.

2.2. Changes in the number of sunken keratosis injuries 2.2.1. Dose effect of triptolide

The results of the evaluation of triptolide administered at a dose of 50 μg or 150 μg per animal are described in Table 1 (number of lesions per mouse and average before and after treatment) and Table 2 (each mouse) The number of lesions and the mean variability before and after treatment).

These results (the average number of injuries per group) are also shown in Figure 1 as a bar graph. Figure 2 is a photograph showing the visual tracking of the skin of representative treated mice of each group over time.

The number of solar keratosis lesions increased systematically between week 17 and week 21 in the untreated areas of all groups. This reflects the new damage that occurred during this period and the microscopic damage at week 17 again developed into visible damage (Fig. 1).

It can be noted that in the control group (treated with vehicle), the number of visible keratosis lesions visible to the naked eye between the 17th week and the 21st week, the treated area, and the untreated area were observed. , increased in equal ways (+72% and +78%, respectively, Table 2). Thus as expected, The agent has no therapeutic effect on solar keratosis damage.

On the other hand, in the two groups treated with triptolide, the number of macroscopic keratotic lesions in the treated area was significantly reduced (50 and 150 μg doses were reduced by 89% and 90%, respectively). It should be noted that triptolide can cause existing lesions to disappear (shown by a systematic reduction in the number of lesions after treatment), as well as no new lesions or microscopic lesions that do not develop into visible damage, and untreated The area is not the same as the control.

The large reduction in the number of lesions after such administration of triptolide reflects its high efficacy in the treatment of solar keratosis. This efficacy appears to be comparable to the performance observed with reference to the product, namely ingenol mebutate and 5-fluorouracil.

Furthermore, at the clinical level, the effect of triptolide is characterized by induction of epidermal erosion in the treated area on the day after treatment, followed by epithelialization of this area. This effect is comparable to the effects observed with reference products, namely ingenol mebutate and 5-fluorouracil. It should be noted that the effects obtained by both of these reference products also appear to reach the dermis and the hypodermis as compared to triptolide treatment, resulting in increased skin toxicity (deep erosion, inflammation), which can interfere with the treatment area. Epithelialization.

2.2.1. Dose effect of triptolide and triptolide PCC

The second study used this solar keratosis model to assess the effect of triptolide (purified form obtained from plants) and triptolide PCC at doses of 10 μg or 50 μg of triptolide per animal. Triptolide is previously defined to contain purified triptolide or tripterygium The composition of the ester PCC is administered. The first group of mice had received 10 μg of triptolide once, the second group of mice had received 50 μg of triptolide once, and the third group of mice (control) received only Carrier. The results obtained are described in Table 3 below.

After the average variation%

These results (the average number of lesions per group) are also shown in Figure 3 as a bar graph. Figure 4 is a photograph showing the visual tracking of the skin of representative treated mice of each group over time (carrier, triptolide 50 μg and triptolide PCC 50 μg).

The number of macroscopic keratosis lesions visible to the naked eye in the area treated between day 116 and day 136 was greatly reduced, with treatment with 50 μg of triptolide and 50 μg of triptolide PCC reduced by 89%, respectively. 85%, and treatment with 10 μg of triptolide and 10 μg of triptolide PCC were reduced by 26% and 27%, respectively. At the same time, the injury in the control mice increased by 30%.

This study confirmed the therapeutic activity of triptolide in the treatment or prevention of solar keratosis in a dose-dependent manner, regardless of the source of triptolide.

2.3. Histological analysis of triptolide effect

The clinical efficacy of triptolide was further confirmed by histology after sacrifice of the animals, and skin sectioning was performed. Indeed, no changes in solar keratosis were observed in the sections taken from the study to terminate the area collected from the treatment. The scar tissue of the skin was observed on the normal epidermis.

A small fraction of epidermal cells have a Ki67-positive marker in the normal epidermis by immunohistochemical labeling of anti-Ki67 antibodies. An increase in the number of proliferating (and thus Ki67-positive) epidermal cells is characteristic of the histology of AK (Acanthosis). Thus, the control mice had a high proportion of Ki67-positive cells. In the epidermal area treated with triptolide, compared with the control group, A large reduction in the number of proliferating cells was observed.

2.4. Determination of triptolide in plasma

The triptolide in the skin and plasma of mice (n=2) treated topically in a closed manner as described above and at a dose of 0.2% (or 400 μg per animal) was determined by mass spectrometry. The amount of product in the plasma was below the lower limit of plasma detection of the method (i.e., below 10 ng/ml), while the treated skin detected approximately 2100 ng/g. Triptolide thus accumulates in the skin and exhibits very low or no transdermal access, which is a very important and advantageous point for topical use.

Example 2: Study of human skin 1. Materials and methods 1.1. Franz static diffusion cell

The kinetics of triptolide and the degree of penetration of human skin are determined by a generally known agreement of the Franz static diffusion cell (OECD Guideline for the Testing of Chemicals: New Guideline 428, Skin Absorption: In Vitro Method ( 13 April 2004)). The Franz groove is used to determine the skin penetration of exogenous substances in the body. In fact, this technique measures the speed at which molecules constituting the active ingredient deposited on the surface of the skin pass through various skin barrier layers. A composition containing the molecule to be studied is deposited in a so-called "donor" chamber of the device. The molecule is then diffused through a membrane, in this case the skin, and collected in a so-called "receptor" chamber. In fact, debris from the skin is deposited in a diffusion trough called the Franz groove. The deep dermis is in contact with the receptor solution, which is considered to replace the interstitial fluid of the skin. A formulation containing the molecules under investigation is deposited on top of the skin. The receptor solution was sampled at regular intervals to determine the molecules that had traversed the degreased skin, and to replace the fresh solution. The amount of material present in the sampled receptor phase can be determined to determine the amount of molecules that have traversed the skin. The skin used in this experiment is the skin of a human donor. The analysis involved liquid/liquid extraction and cortisol as an internal standard, which was then analyzed by liquid chromatography coupled tandem mass spectrometry (LC-MS/MS).

1.2. Composition and application agreement

The compositions used were as described in Table 4 below. Placebo formulations served as controls.

2. Results

The results of the test for the distribution of the transdermal triptolide are described in Table 5 below, which shows the distribution of triptolide (expressed as the percentage of application) in each chamber:

As expected, triptolide was not detected after administration of the placebo formulation.

After application of Composition A, the average total recovery of triptolide was about 85%. This result is universal and acceptable in the state of the art, first considering the use of non-radiolabeled triptolide and, secondly, the cumulative error margin of each chamber.

In summary, the results show that the degree of absorption of triptolide after 24 hours of application is very low (about 1.1% of the applied amount is considered to be bioavailable). This amount corresponds to the amount found in the skin (epidermal + dermis) and the receptor fluid (RF). Triptolide is known to be very active and is also toxic in large quantities. The result of this low level of systemic throughput is unexpected and unexpected, and makes it possible to consider topical application of triptolide without risk to humans.

Claims (12)

Use of a compound selected from the group consisting of triptolide, derivatives thereof, and pharmaceutically acceptable salts of triptolide and its derivatives for topical treatment and/or prevention of tumors or cancers of the skin Pre-pathology. The use of the compound of claim 1, wherein the triptolide derivative is selected from the group consisting of triptonide, tripdiolide, triptolide Triptolidenol, 16-hydroxytriptolide, triptriolide, 12-epitriptriolide, 14-epitriprin (14-epitriptolide), tripoxalide and derivative PG-490-88. The use of the compound of claim 1 or 2, wherein the triptolide derivative is triptolide or triptolide. Use of a topical composition comprising one or more compounds according to any one of claims 1 to 3 and at least one pharmaceutically acceptable excipient for the treatment and/or prevention of tumors or precancerous skin pathology. The use of the topical composition of claim 4, wherein the compound is triptolide or a pharmaceutically acceptable salt thereof. The use of the topical composition of claim 4 or 5, wherein the compound is present in an extract obtained from a family of the family Celastraceae . The use of the partial composition of any one of claims 4 to 6, wherein The concentration of the compound is between 0.0002% by weight and 2% by weight based on the weight of the final composition. The use of the topical composition of any one of claims 4 to 7, wherein the compound is between 1 μg/cm 2 (μg/cm ² ) and 1.5 mg/cm 2 (mg/cm ² ) of skin. The dose is administered. The use of a compound according to any one of claims 1 to 3, wherein the tumor or precancerous pathology of the skin is selected from ultraviolet light in the skin, or the use of the topical composition according to any one of claims 4 to 8. Tumor and precancerous pathology, metastasis, and T-cell lymphoma of the skin. The use of the compound of claim 9 or a topical composition, wherein the ultraviolet-induced tumor or precancerous pathology of the skin is selected from the group consisting of solar keratosis, basal cell carcinoma, melanoma, and Squamous cell carcinoma. The use of the compound of claim 10 or a topical composition, wherein the ultraviolet-induced tumor of the skin or the precancerous pathology is solar keratosis. The use of the compound of claim 10 or a topical composition, wherein the ultraviolet-induced tumor or precancerous pathology of the skin is derived from squamous cell carcinoma of solar keratosis lesions.