MXPA96004956A - Porfocianin and porfirinas extended with - Google Patents

Abstract

The present invention relates to a compound of the formula I: and the metallated forms and salts thereof, characterized in that ne is an integer from 1 to 4, and wherein each Pi is independently a pyrrole residue, wherein each Z is independently a covalent bond, or is a meso bridge forming group, a N-meso bridge forming group, a CC bond of the formula Rid-C = CR

Description

PORFOCIANIN AND PORFIRINAS EXTENDED WITH CMC Related Application This application is a continuation in part of Serial No. 08 / 077,789 issued on June 15, 1993, which is a continuation in part of Serial No. 07 / 968,966, issued on October 30, 1992; descriptions of which are incorporated herein by reference.

Field of the Invention The invention relates to porphyrins extended with CNC, such as porfocyanines, to methods for their preparation and the use of these compounds to act as a mediator in the detection or destruction of target cells or tissues by the irradiation of light. In addition, the invention relates to the use of the compounds of the invention in methods of radioforming imaging and magnetic resonance imaging.

Background Technique Although a considerable effort has been devoted REF: 23311 to the synthesis and study of porphyrins and other tetrapyrrolic macrocycles, less is known about the larger, aromatic pyrrole-containing systems, the so-called "extended porphyrins". Such systems, by virtue of containing a greater number of ft electrons, additional coordination heteroatoms and a larger central link core, may offer advantages over porphyrins. Many of the extended porphyrin systems contain more than 4 pyrrole rings. The synthesis of safirin from tripirran dicarboxylic acid and bipyrroldicarboxaldehyde to produce a pentapyrrole compound was reported several decades ago by Woodward, RB, Aromaticity Conference, Scheffield, England, 1996. See also, Broadhurst et al., J Chem Soc Perkins Trans ( 1972) j_: 2111 and Bauer et al., J Am Chem Soc (1983) 1_05: 6? 29). The synthesis of esmeral-dirina from bipirroldicarboxaldehyde and pyrroldipyrrometan dicarboxylic acid was reported in 1970 by M.M King. (Ph.D. Dissertation, Harvard University, Cambridge, MA). The uranyl complex of superftalocyanin is another historically important macrocyclic pentapyrrolic compound. This compound was prepared by the direct pattern condensation of dicyanobenzene with uranyl dichloride, however, the free base is unstable (Day et al., J Am Chem Soc (1975) 97: 4-519). The demetalation results in the contraction of the ring to form phthalocyanine (Marks, T. J. and D.R. Stojakovic J Am Chem Soc (1978) 100: 1695). Gossauer synthesized the first hexapyrin by condensing a bis-alpha-tripyrrane with a tripyrran dialdehyde, followed by oxidation (Bull Soc Chim Belg (1983) 92: 793). Of the six methane bridges present in hexaphyrin, two have the E (Id.) Configuration. Charriere reported that hexaphyrin forms bimetallic complexes with various transition metals (1987, Thesis, University of Friborg, Suisse). Another hexapyrrolic system, rubyrin, has recently been synthesized and structurally characterized (Sessler et al., Angew Chem Int Ed Engl (1991a) 30: 977). Porphyrins or platyrins vinylogas (related chemical type, but different because it has one or more vinylene bridges between the functional atoms in the organic molecule) are another important class of macrocycles containing pyrrole, first described by R.A. Berger and E. LeGoff (TetraLett (1978) 4 .: .2; see also, LeGoff, E. and OG Weaver J Org Chem (1977) 52: 711; and Franck and collaborators Proc SPIE Int Soc Opt Eng, Ser 5 (1988) 997: 107).

These compounds were generally synthesized by reacting a dipyrromethane with a dipyrromethane substituted with acrylaldehyde (Beckman et al. Angew Chem Int Ed Engl (1990) 29: 1395). Extended bisvinyl porphyrins were further extended to tetravinyl porphyrins in which all four meso bridges become enlarged. Tetravinyl porphyrins are made by an acid-catalyzed autoconditioning of the N-protected pyrrole-substituted allyl alcohol. Tetravinyl porphyrins have a very strong Soret-like band change of more than 150 nm from that of normal porphyrins (Gosmann, M. and B. Franck Angew Chem Int. Ed Engl (1986) 25: 1100; Knübel, G. and B. Franck Angew Chem Int Ed Engl (1988) 27: 1170). In addition, bisvinyl porphylene synthesis has been recently restated (Jux et al. Angew Chem Int Ed Engl (1990) 29: 1385; Vogel et al. Angew Chem Int Ed Engl (1990) 29: 1387). The Schiff-based compounds represented by the texaphyrins comprise other class of macro-cycles containing pyrrole (Sessler et al. J Org Chem (1987) 52: 4-394-; Sessler et al. J Am Chem Soc (1988) O: 558β ) Sessler, JL and collaborators Acc CHem Res (1994) 27: 43-50. Texaphyrins are synthesized by the acid catalyzed condensation of tripyrran dialdehyde with o-phenylenediamine. Several texaphyrins have been prepared using similar strategies (Sessler et al. (1991) Abstract of the 201st Nati Soc Mtg, Inorganic Division; et al. Inorg Chem (1992) 28: 529). The use of porphyrins, combined with irradiation, for the detection and treatment of malignant cells has, for this time, some considerable history. (See, for example, Porphyrin Photosensitization (Kessel, D. and collaborators, eds Plenum Press, 1983). Certain porphyrins appear "naturally" capable of localizing malignant cells. When irradiated, porphyrins have two properties that make them useful. First, when irradiated with ultraviolet or visible light, these can be fluorescent, and thus be useful in diagnostic methods related to the detection of malignancy (see, for example, Kessel et al., Supra; Gregory, HB Jr et al., Ann Surg (1968) 167: 827-829 ). In addition, when irradiated with light, certain porphyrins exhibit a cytotoxic effect on the cells in which they are located (see, for example, Diamond, I. et al, Lancet (1972) 2 ^ 1175-1177; Dougherty, TJ and collaborators, Cancer Research (1978) 38: 2628-2635; Dougherty, T.J. and collaborators, The Science of Photo Medicine 625-638 (J. D. Regan &J.A. Parrish, eds., 1982); Dougherty, T.J. and collaborators, Cancer; Principles and Practice of Oncology 1836-1844 (V.T. DeVita Jr et al., Eds., 1982). It is true that extended porphyrins such as safirin, texaphyrin and vinyl porphyrins have unique long wavelength and individual oxygen producing properties, which also make them attractive as potential photosensitizers for use in phototherapy of the tumor (Maiya et al. J Phys Chem (1990) 9 ^: 3597; Sessler et al SPIE Soc (1991) 1426: 318; Franck et al., Supra). Porphyrins, such as hemato-porphyrin derivatives, monohydrobenzoporphyrin (BPDs) and sodium porfimer have been conjugated to target cell-specific immunoglobulins to improve their ability to harbor the desired cells or tissue. A problem dependent on the practice of photodynamic therapy is that the wavelength for the irradiation required to activate certain porphyrins is in the range of 630 nm, wavelength which is also easily absorbed by other porphyrins and natural chromophores normally present in the blood and other tissues. Therefore, the depth of effective treatment has been limited to a few millimeters as it blocks the effects of natural chromophores that absorb light such as hemoglobin. When a greater penetration is desired, it is desirable to administer compounds to act as mediators of the irradiation effects that can be exited at long wavelengths such as BPDs, thus avoiding the blocking effects of natural chromophores present throughout the organism of the subject. In addition to phototherapy, extended porphyrins are useful in magnetic resonance imaging (MRI). MRI is a non-ionizing, non-aggressive method that allows normal and abnormal tissue to be observed and recognized in the early stages of development. At this time, MRI has a significant disadvantage, in that the degree of signal increase for diseased versus normal tissues is often insufficient to allow this method to be used in many clinical situations. To overcome this problem, contrast reagents have been developed for MRI. Paramagnetic metal complexes, such as those derived from gadolinium (III) (Gd) have recently demonstrated particular efficiency in clinical trials. To date, the coordination of gadolinium in contrast agents for MRI has been achieved using carboxylate ligands that include porphyrin, but more successfully with texfifirins. (See, for example, Lauffer, RB Chem Rev (1987) 87: 901; Kornguth et al. J Neurosurg (1987) 66: 898; Koenig et al. Invest Radiol (1986) 21: 697; chacheris et al. Inorg Chem (1987) 26: 958; Loncin et al. Inorg Chem (1986) 25_: 2646; Chang, C.A. and V.C. Sekhar Inorg Chem (1987) 26: 1981). Sessler et al report that texaphyrin forms an extremely stable Gd (III) complex in vitro (Sessler et al. Inorg Chem (1989) 28: 3390). Gd (III) does not form stable complexes with normal porphyrins. In addition to Gd (III), texaphyrin has been reported to form complexes with a variety of transition metals such as Cd and Eu (Sessler, J.L. and A.K. Burrell Top Cur Chem (1992) 161: 177).

Description of the invention The invention provides novel light absorbing compounds suitable for use in detecting and / or treating target tissues, cells and pathogens. The compounds of the invention can be used in photodynamic therapy and in diagnosis in a manner analogous to that in which porphyrins, phthalocyanines, BPDs, and related compounds can be used. If desired, the compounds of the invention can be administered in relatively low doses due to their ability to absorb radiation in an energy range outside that normally absorbed by the components present in high concentration in the blood or other tissues, in particular, the porphyrin residues normally associated with hemoglobin and ioglobin. This is advantageous when the penetration of tissues by light is required. In some cases, where surface treatment is sufficient, shorter wavelengths may be used. These compounds are preferentially retained in tissues and target cells as compared to tissues and non-target cells, although this property may not be necessary for effective treatment. See, for example, US Patent Serial No. 07 / 948,113 issued September 21, 1992 and 07 / 979,546 issued November 20, 1992, the contents of which are incorporated herein by reference. Thus, in one aspect, the invention is directed to extended porphyrins with CNC of the formula PrZr (Pr2l) .- Pl-Z, I (1) where n = 1-4; wherein each P. is independently a pyrrole residue of the formula H wherein each R. and R- is independently a substituent without interference, and wherein each Z. is independently a covalent bond; or is a bridge group meso of the formula a group of N-meso bridge of the formula a CC link of the formula R R R? and id R? e - * = c- c = or is a CNCCNC link of the formula where each R. > R- J > R, - "> R f and R- is independently a substituent without interference; or is a CNC link of the formula - CH = N-CH = + a * = CH-N = CH- where at least one of Z. is the CNC link. In the compounds of the invention at least one Z. is the CNC link. The compounds of the invention also include the metallated forms and the salts of the compound of the formula (1). The prototype of this group of "porforinas extended with CNC" is porfocianina, as it is described later. In other aspects, the invention is directed to pharmaceutical and diagnostic compositions containing the compounds of the invention, for conjugation wherein the compounds of the invention are covalently linked to specific target agents, such as immunoglobulins and for the use of these compounds or conjugates in photodynamic therapy and in diagnosis. In another aspect, the invention is directed to the forms of the compounds of the invention suitable for use as contrast agents for the radioforation of images and magnetic resonance imaging, as well as methods for using them. Another aspect relates to the synthesis of the compounds of the invention.

Brief Description of the Drawings Figure 1 shows the emission spectrum of octaethylporfocyanine in the free base form.

Figure 2 shows the octaethyloprofocyanine emission spectrum in the acid addition salt form.

Figure 3 shows some preferred embodiments of the compounds of the invention.

Ways of Carrying Out the Invention The compounds of the invention, herein referred to as "CNC-extended porphyrins", contain at least 3 pyrrole cores bonded through covalent bonds, or through optionally substituted conventional meso bonds found naturally in porphyrins, or through N-bonds. meso, or through optionally substituted CC bonds as found in the porfacenes or through optionally substituted CNCCNC bonds analogous to the o-phenylene diamine linkages found in texafins, or by CNC links, but where at least one link is a CNC link of the formula C H = N- C H = = C H- N = C Members of a specific class that belongs to this group are designated by phocyanins. These are represented by the formula the various R substituents are those which do not interfere with the basic porfocyanine nuclei in their ability to absorb light of appropriate wavelengths and to achieve at least one desired effect, such as acting as a mediator in the destruction of tissues and target cells in a context of otodynamic therapy, serve as a contrast agent for MRI, and the like. The nature of suitable R substituents, which will not interfere, is discussed further below. However, it can be established that a wide variety of substituents can be employed without interfering with the utility of these compounds. The proper selection of these substituents without interference will depend on the specific use proposed. For example, if it is proposed for in vivo use, substituents should not be toxic. When used in vitro, especially if the possibility for the removal of the compounds of the invention from the appropriately treated material is available, this may not be of importance. These or the ordinary expert will understand that the parameters that restrict the selection of each In addition to the compounds of the formula (1a), other extended porphyrins with CNC are included in the scope of the invention. In a preferred embodiment, where n = 2, formula (1) can be represented by L P 7 L z- where each P. and Z. is as defined above. Illustrative of these compounds -CNC- where n = 2, are the compounds of the formulas (lb) - (1e): or, generically, As seen in these illustrative formulas, at least one bridge between the four pyrrole cores must be a CNC link. Otherwise, the linkage may be a covalent bond or the indirect, meso, conventional, discrete methane bond optionally substituted by Rí.c, 'or the other links described above. The restoring arrangement of tf-bonds in the CNC porphyrins of the invention will be dependent on the selection of the link, as understood by those experts '^ .0 ordinary in organic chemistry. For example, replacing an odd number of meso links with CNC converts an aromatic system to a non-aromatic one, even a stable one; the replacement of even a number retains aromaticity. 15 Additional modalities include those where n = 1: or where n = 3 or where n = 4: Particularly preferred are those modalities shown in Figure 3. With respect to the substituents R.sub.a, R.sub.i, Ric.Rid.Rie.Rif and Rig in general, each R.sub.in is independently halo, nitro, cyano, NR '", SR', OR ', SOR', S02R ', S00R', C0NR'2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, or optionally substituted aralkyl. The substituent R 'represents hydrogen or (1-6C) alkyl.

The amino, sulfhydryl and hydroxyl substituents can also be acylated (1-6C). Suitable substituents for optional substitutions include those specified above, ie, halo, nitro, cyano, NR'2 > SR ', OR' and the like. Of course, the aromatic nuclei can also be substituted by alkyl, alkenyl or alkynyl groups. As used herein, alkyl, alkenyl and alkynyl are conventionally defined as hydroxycarbyl substituents containing 1-6C, are either saturated or unsaturated, and are straight chain, branched or cyclic portions. In this manner, the alkyl would include methyl, tertiary butyl, cyclohexyl, n-hexyl and the like; the alkenyl would include these major carbon columns with one or more double bonds; and the alkynyl would include similar carbon sturctures with one or more triple bonds. Arylalkyl (5-18C) refers to a substituent bonded to the porphyrin nuclei extended with CNC through an alkylene group, wherein alkylene is defined in a manner corresponding to the definition of (1-6C) alkyl. By "aryl" is meant an aromatic portion of 4-12C, optionally containing one or more heteroatoms. In this manner, suitable aryl groups include phenyl, naphthyl, pyridyl, pyrimidyl, quinolyl and the like. Where the substituents contain carboxylic acid or amino substitutions, the relevant salts are also included within the scope of the compounds of the invention. The -COOH salts can be derived from inorganic or organic bases, including non-toxic, pharmaceutically acceptable inorganic and organic bases. Suitable inorganic bases include sodium, potassium, lithium, ammonium, calcium and magnesium, hydroxides and the like. Particularly preferred are the potassium and sodium salts. Organic non-toxic, pharmaceutically acceptable bases include primary, secondary, tertiary and quaternary amines including cyclic amines and ion exchange / basic resins. Examples include isopropylamine, trimethylamine, ethanolamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, glucose-mine, theobromine, purines, piperazine, polyamine and the like resins. Any amino group contained in the substituents may also be present in the form of their acid addition salts. In addition, porphyrin nuclei extended with CNC themselves can be present in the form of an acid addition salt.

These salts are formed of inorganic acids such as hydrochloric, sulfuric and phosphoric acid or with organic acids such as acetic, oxalic, benzoic acid and the like. The conversion of the compounds of the invention, free acid or free base to the corresponding salt forms and vice versa is conducted by methods well understood in the art. the above-mentioned moieties of R1.a and J Ri., b include hydrogen, optionally substituted alkyl, optionally substituted aryl and optionally substituted arylalkyl. A particularly preferred substituent is carboxyl. More preferred are hydrogen and alkyl. In addition, while each R. and R., independently is determined, the symmetry in the selection of these substituents facilitates the preparation of the compounds since the number of components in the mixture of the resulting product is reduced. While chromatographic separation techniques are adequate to allow an arbitrary number of components to be separated from the product mixture, yields are increased when the number of possible products is reduced. Thus, particularly preferred are the compounds of the invention wherein all R X.3. they are the same and all R. ~, u are the same, especially where all R? .a and RI., D are the same.

Another preferred embodiment is that wherein the compound of the formula 1 (a), R1 & , R ^, R, and R "are the same and wherein" ^ 2b 'R3a' and R3b are the same as each other. although different from R1, R.,, R. and R, -.

Particularly preferred embodiments of R 1. C-Rl.g include H and optionally substituted aromatic (aryl) substituents. Particularly preferred embodiments of R. are H and optionally substituted phenyl. Illustrative preferred embodiments of the compounds of the invention where n = 1 are: 1. Z- 1. is CNC, Z is it meso where R c is H, Z is a covalent bond, R- and R2b are H, and R-? B, R, R ~ and Rr., Are methyl; 2. Z., is CNC, Z is meso where R is phenyl, Z is a covalent bond, R-? A, Rpn 'R3a and R ,, are (CH2) 2C00H, R1b is methyl and R2b is H; 3. Z1 is CNC, Z2 is CNCCNC where R2f J and R2g are H, ZJ, ¡CC where Rjjc, ly "RJ-e are H, and where R1a and R2b are H, and R1b, R2a, R ^ a and R ^ b are methyl, 4. Z1 is CNC , Z2 is CNCCNC wherein R2f and R? Are phenyl, Z ^ is CC where R_, and R-, are phenyl, R1a 'R2a' R3a and R3b are (CH2) 2C00H, ^ is methyl and R2b is H; . Z1 is CNC, Z2 is CNCCNC wherein R2f is phenyl and R is H, and Z is a CC bond where R ~ is phenyl and R ~ is H and where R? , R_ and R, are phenyl and R ,,, R2b and Rb are H. The preferred embodiments where n = 2 are: 1 Z1 is CNC, Z2 is N-meso, Z ~ is meso where R, is methyl, and Z, is CNC and where R .. and R2b are H and R1b, R2a, R3 &, R ^ b, R ^ and R ^ b are methyl; 2. Z., is CNC, Z2 is N-meso, Z ~ is meso where R, is phenyl and Z, is CNC and where R-? A > Rpa ' R3a 'R3b and R4a are (CH ^ COOH and R1b, R2fe and R ^ are H; 3. Z1 is CNC, Z? Is N-meso, Z ~ is meso where R3 ~ c is H,' and ^ Z, is CNC and where R.1 a, R «a.R3, a" and R ^ a are phenyl and R1b, R2b, R ^ and R ^ b are ethyl, 4. Z1 is CNC, Z2 is CNCCNC where R2f and J R2g are both phenyl, 'Z3q is a CC bond where R-j and R are H, and Z, is a covalent bond and where R1a and R2b are H and R1b 'R2a' R3a 'R3b' R4a and R4b are methyl; 5. Z1 is CNC, Z2 is CNCCNC where R2f and R "are H, or Z, is a CC bond where R ~ d is ethyl and R-, is H, and Z, is meso where R, is ethyl and in J 3? 'J 4 4c where R1a, R2a, 3 & , R3b 7 R ^ a are (CH2) 2C00H and R1b, R2b R4b s on HJ 6. Z1 is CNC, Z2 is CNCCNC where R2f is ethyl and J R2g is H, Z3 is meso where R3-c is phenyl and Z. is a CC bond where R_, and R_ are methyl and where R1la, R < a, R_.?a, and R-y + a_ are phenyl and R1IvD. -D > 3h and R / b are e "kilo In the examples where n = 3, the preferred compounds are: 1. Z-, is CNC, Z2 is meso where Rp is methyl, Z- is meso where R ^ is methyl , Z, is a 3 3c '4 covalent bond and Z is CNC and where R1, R, R-, ^ a and 5a are methyl and R1b, R2b, R3b, R ^ and R5b are H; 2. Z1 is CNC, Z2 is N-meso, Z_ is meso where R_ is phenyl, Z, is CNC and Z c is CC where R, and R5e are phenyl, and wherein R1a> R1fe, R ^, R ^, H ^ and R ^ are (CH2) 2C00H and 2a 'R2b' R4a and Rb are metily> 3. Z1 is CNC, Z2 is N-meso, Z is meso where R-, is H, Z, is N- meso, and Zr is CNC and where R1a, 3a 5a are phenyl, R1b, R ^ and R5fe are H and R2a, R2b 'R4a and R4b are methyl ° »» - is CNC, Z2 is meso where R2 is methyl, Z_ is CNCCNC where R ~ "and R., are phenyl, Z is N-meso, Zr is a CC link where and R c are H, and where R_¡ & and R2b are H and R ^, R2a > R3 &, R3b > R. and R., are ethyl; 4a J 4b 5. Z1 is CNC, Z2 is CNCCNC wherein R2f and R are H, Z is a CC bond where R J is ethyl < g 3 3d and R30e is H, Z4, is meso where R4, c is ethyl J and Z 5r is a covalent bond, and where R1, R, R-,, R ^, and R4a are (CH2) 2C00H and R1fe, R2fc, R ^ b, R ^ and R ^ are H; 6. Z1 is CNC, Z2 is CNCCNC wherein R2f is ethyl and R2 is H, Z-, is meso where R ^ is phenyl, Z, is a CC link where R ,, and R, are methyl and 4 4d J 4? "^ Z ^ is CNC, and wherein R1ft, R2a, R3a, R ^ a and R5 & are methyl and R-? b, ph 'R3b' 4b and R5b are e "kil; 7. Z .. is CNC, Z2 is a CC bond where 2d and R2e are phen; i-l0> 2 is CNCCNC in where R ^ f and R are H, Z, is N-meso and Z is meso where Rr is methyl and in giving R3a 'R3b' R5a and R5b are (CH ^ COOIT and R2a, R2b, R, and R, b are methyl; 8. Z1 is CNC, Zp is a covalent bond, Z_ is meso where R-, is phenyl, Z, is CNC and Zr is a CC bond where Rrj and r are methyl and where R1a 'R3a and R5a are phenyl > Rlb 'R3b and R5b are H and R2a' R2b 'R4a and R4b SOp pe il0' For compounds where n = 4, preferred embodiments include: 1. Z .. and Z¿ are CNC, Z2 and Zr are CC links where all R2d, R2e, Rrd and Rre they are phenyl, and Z3 and ^ are meso where R3c and R ^ are ethyl, and where R1a 'R2a' R3a 'R4a' R5a and R6a are i-propyl and R1fe, R2b, R3b, Rb, R5b and R6b are (CH2) 2C00H; 2. Z1 is CNC, Z2 and Z, are N-meso, Z3 and Z? are covalent bonds and Z¿ is meso where R¿ is H and R1a 'R2a' R3a 'R4a' R5a and R6a are phenyl and R1b 'R2b' R3b 'R4b * R5b and R6b are methyl ° - The possibility of obtaining porphyrins extended with CNC with a multiplicity of substituents has the advantage that the properties of porphyrin nuclei extended with CNC from the center can be modified by the selection of such substituents. For example, the solubility of the compounds can be increased by employing substituents with polar groups. By altering the symmetry of the arrangement of these groups, the compounds can be made amphiphilic, that is, the compound will have a configured distribution of charges + and -. This property is useful for solving problems of biodistribution - membrane transition and the like. In addition, the amounts of light absorption from the central core can be modified to some degree by the conjugated t bonds in the substituents that give the usual auxochromic change.

Metalation / Adding Radio Isotopes The CNC-extended porphyrin compounds of the present invention include those where the extended porphyrin with CNC itself is coupled with a radioisotope for image radiophoresis (scintigraphic image formation) or with certain metals for use as ions, an agent of contrast of magnetic resonance imaging, or simply as a convenient form of the compound. Examples of radioisotopes that would be useful labels include Iodine-123, Iodine-131, Technetium-99m, Indium-111 and gallium-67. Examples of metals that would be suitable as contrast agents for MRI include paramagnetic ions of elements such as Gd, Mn, Eu, Dy, Pr, Pa, Cr, Co, Fe, Cu, Ni, Ti, and V, in the form Preferred Gd and Mn.

Conjugates of the Compounds of the Invention For use in some contexts, it is useful to bind the compounds of the invention to a specific target portion such as an immunoglobulin or a ligand for a receptor. The ability of the compounds of the invention to lodge in diseased tissues and cells such as tumors, if desired, can be increased by coupling the compound to a portion that binds epitopes or receptors located on the surface of such tissues or cells. objective. In this manner, the specific target portions within the present invention include ligands such as steroids, such as estrogen and testosterone and derivatives thereof, peptides comprising ligands for T cell receptors, saccharides, such as mannose for monocytes and macrophages to have receptors, and H2 agonists. The specific target portions can also be immunospecific components. SSTs can be derived from polyclonal or monoclonal antibody preparations and can contain whole antibodies or immunologically reactive fragments of these antibodies such as fragments (F (ab ') p, Fab or Fab' .The use of such immunologically reactive fragments as substituents for Complete antibodies are well known in the art, See, for example, Spiegelberg, HL, Immunoassays in the Clinical Laboratory (1978) 3: 1-23. Polyclonal antisera are prepared in conventional ways by injecting a suitable mammal with antigen to the antibody that is desired, assaying the level of the antibody in serum against the antigen, and preparing the antisera when the titers are high. Monoclonal antibody preparations can also be prepared in a conventional manner such as by the method of Koehier and Milstein using peripheral blood lymphocytes or spleen cells from immunized animals and immortalize these cells either by viral infection, by fusion with ielomes, or by other conventional procedures, and select the production of the desired antibodies by isolated colonies. Formation of the fragments of any monoclonal or polyclonal preparation is carried out by conventional means as described by Spiegelberg, H.L., supra. Particularly useful antibodies exemplified herein, include the preparation of the CAMAL-1 monoclonal antibody, which can be prepared as described by Malcolm et al. (Ex Hematol (1984) 1.2: 539-547); polyclonal and monoclonal preparations of anti-M1 antibody as described by Mew et al. J Immunol (1983) 130: 1473-1477 and antibody B16G which is prepared as described by Maier et al. J Immunol (1983) 131: 1843 and Steele and collaborators Cell Immunol (1984) 90: 303. The above list is empirical and not limiting; once the target tissue is known, an antibody specific for this tissue can be prepared by conventional means. Therefore, the invention is applicable to effect toxicity against any desired objective.

The coupling of the specific target portion to a CNC expanded porphyrin of the present invention can be effected by any convenient means known in the art, depending on the nature of the substituents on the porphyrin portions extended with CDC. For example, if at least one R. contains a carboxylic group, a covalent bond can be made to an amino-containing SST using a dehydrating agent such as carbo-diimide. A particularly preferred method of covalently linking a porphyrin extended with CNC to the specific target portion is treatment with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCI) in the presence of a reaction medium consisting essentially of dimethyl sulfoxide (DMSO). A preparation is then illustrated using this preferred method. Other dehydration agents such as dicyclohexylcarbo-diimide or diethyl sarbodiimide can also be used, as well as conventional aqueous and partially aqueous media. The active portions of the conjugate can also be coupled via linker compounds which are bifunctional, and are capable of covalently bonding each of the two active components. A large variety of these linkers is commercially available, and a typical list would include those found, for example, in the catalog of the Pierce Chemical Co., Rockford, IL. These linkers are either homo- or heterobifunctional portions and include functionalities capable of forming disulfides, amides, hydrazones and a wide variety of other linkages. Other linkers include polymers such as polyamines, polyethers, polyamine alcohols, derivatized to the components by means of ketones, acids, aldehydes, isocyanates, or a variety of other groups. The techniques employed in coupling the active portions of the conjugate include any conventional means and the method for coupling does not form part of the invention.

Use of the Compounds of the Invention in PPT and Other Applications The compounds of the invention can be used in photodynamic therapy protocols analogous to those known for sodium porphyria and BPDs. In this way, the compounds can be used to destroy or impair the functioning of unwanted cells or tissues or for inactive pathogens such as bacteria and viruses.

Examples of target cells and tissues for which PDT is useful include, but are not limited to, tumors, including blood tumors, malignant bone marrow, virally infected blood or bone marrow cells, cells and dysplastic tissues, sites of inflammation or infection, hyperproliferative tissue such as psoriatic plaque or papilloma-virus lesions (wart) or lesions of neointimal hyperplasia, hypervascularization such as congenital capillary hemangioma and hema-giomas, atherosclerotic plaque, hair follicles, free viruses, bacteria , - protozoa or other pathogenic parasites. Pathogens that can be adversely affected by PDT include certain viruses, bacteria, protozoa and other pathogenic parasites. Viruses, for example, include developed viruses such as human cyto-megalovirus, Epstein-Barr virus, Marek's disease herpes virus, human herpes simplex virus, varicella-zoster virus, members of the Poxviridae family, members of the Hepadnaviridae family such as human hepatitis A virus (HAV), human hepatitis B virus (HBV) and virus without A, without hepatitis B, including human hepatitis C virus, members of the Orthomyxoviridae family such as influenza virus or influenza types A, B, and C, members of the Retroviviridae family such as human T cell leukemia virus, human immunodeficiency virus, and members of the Flaviviridae family such as tick-borne encephalitis virus or fever virus yellow. Illustrative parasites include Plasmodium malariae, P. falciparum, P. ovale, P. vivax and Trypanosoma cruzi. Bacteria include Bacillus subtilis, Streptococcus faecalis, Pseudomonas spp., Mycobacterium spp. and other opportunistic organisms that can be treated by photodynamic activation. The manner of use of the compounds of the invention in these contexts follows the procedures that are established in the art. The selection of wavelength absorption for extended porphyrin with CNC used in a particular technique can be induced by the nature of the use. For example, where the target tissue is in a context with a number of interfering materials such as those found in the blood, it is advantageous to use extended CNC porphyrins that have wavelength absorptions undivided with these contaminants. On the other hand, for the treatment of superficial target tissues, this consideration is not relevant. In this way, the presence or absence of alternative compounds capable of absorbing light in the environment,. , to be irradiated with light, is taken into consideration when selecting an appropriate member of the extended porphyrin class with CNC of the invention and in the wavelength selection. In addition to their use in photodynamic therapy, the compounds of the invention can be used for diagnosis in a manner analogous to that for porphyrins per se. The extended porphyrins with CNC, similar 'porphyrin wings themselves, are able to have, fluorescence when stimulated with light of an appropriate wavelength. As the porphyrins extended with CNC, similar to the porphyrins themselves, they will harbor certain target tissues, such as atherosclerotic plaques, tumors and the like. If appropriate, the compounds of the invention can be provided with specific target portions to ensure this "sheltered". The compounds of the invention can thus be allowed to accumulate in such tissues or target cells if they are present, and these tissues or cells can be discover or locate by detecting fluorescence. In addition, the compounds of the invention can include radioisotopes either as covalently bound substituents or in the metallated complexes so that the compounds of the invention can be to locate by the radioformation of images or scintigraphic. Certain metal ions also provide the compounds of the invention with the ability to serve as contrast agents for MRI. These contrast agents are conventionally used when obtaining scans by MRI with increased interest.

Preparation of the Compounds of the Invention A number of strategies can be used to perform the CNC link of the invention. A single CNC link can be formed, for example, by reacting a substituted pyrrole at the 2-position with a carboxylic acid moiety with a pyrrole substituted at the 2-position with aminomethyl. This general method for forming the CNC link is shown in Reaction Scheme 1.

Reaction Scheme 1 As shown, conventional amide formation is followed by dehydration to form the CNC link of the invention. The CNC link is stabilized by its inclusion in the resonance system of the extended porphyrin nuclei. An additional approach is illustrated by Reaction Scheme 2, which effects the condensation bond of two cyano groups, each link to position 2 of an attached pyrrol.

M H j C H Reaction Scheme 2 As shown in Reaction Scheme 2, of dipyrrole substituted with cyano in this case, it is first reduced with lithium aluminum hydride and then treated with oxygen to form the resulting bond. The reduced compound (3) is partially ionized and the ionized form is combined with the compound of the formula (3) in the presence of 02 to give the product (1). An alternative to Reaction Scheme 2 is shown in Reaction Scheme 3. Instead of treating the reduced dicyano compound with lithium aluminum hydride with oxygen; DDQ is used in methylene chloride and THF.

Reaction Scheme 3 It will be apparent from Reaction Scheme 3 that preferably starting with the bisnitrile, the starting materials could also comprise the dipyrromethane derived from bisaminomethyl shown as the formula (3). This starting material is especially used advantageously when obtaining asymmetric forms of the porfocyanines of the formula (1a). This is illustrated in Reaction Scheme 3 ', which illustrates the condensation of two different "halves" of the porfocyanine nuclei to obtain the desired asymmetric product. Of course, the two symmetrical products are also formed, but these are easily separated from the desired asymmetric product using normal chromatographic techniques.

Reaction Scheme 3 In another variant of Reaction Scheme 3 ', preferably to condense two diaminomethyldipyrromethanes as shown, bisaldehyde (which can be obtained as an intermediate in Reaction Scheme 5 below) is added. This facilitates the production of asymmetric forms of porfocyanines in which the nature of R- | a > Rib 'R4a and R4b ^ as ^ corao R? C) may be different from 2a > R2b 'a' b and R3c * ^ In this manner, the substituents in the compound of the formula (3) shown in Reaction Scheme 3 may be different from those of the dialdehyde and an individual asymmetric product is obtained. In the alternative, a dialdehyde form of the dipyrrole intermediate can be condensed directly using ammonia in ethanol and water as shown in Reaction Scheme 4.

H Reaction Scheme 4 The portion of dipyrrole that contains two cyano groups for condensation in the Reaction Schemes 2 or 3, or dialdehyde for condensation in Reaction Scheme 4, can be obtained as shown in Reaction Scheme 5. 1 H-NOH, NaOAc + CH20 2 Ac20 Reaction Scheme 5 As shown, dipyrrole is formed by the condensation of the 2-benzoyl carbonyl-4-acetoxylmethyl pyrrole molecules. The resulting dipyrrole is reduced to the corresponding dialdehyde and converted to the cyano derivative using normal reagents. Finally, Reaction Scheme 6 shows a further alternative to obtain the dicyan intermediate represented by formula (2).

The dicyano compound can then be converted to the appropriate porphocyanine using the approaches set forth above in Reaction Schemes 2 or 3.

Reaction Scheme 6 In the initial step of Reaction Scheme 6, benzaldehyde optionally substituted with 1-5 substituents independently selected to form the meso bridge between the two condensed pyrrole nuclei is used. However, other aldehydes, including formaldehyde and simple alkyl aldehydes such as acetaldehyde, butyraldehyde, hexalaldehyde and the like can also be used. In addition, other aryl aldehydes such as 1-naphthylaldehyde, can also be used. In the second step, the chlorosulfonyl isocyanate (C1S02N = C = 0; CSI) in a polar aprotic solvent, such as DMF, derivatizes the pyrrole nuclei so that they contain the desired cyano groups at positions 5. The specific components shown in Reaction Scheme 6 are proposed to be non-limiting illustrative. In more pertinent details regarding Reaction Scheme 5, dicyanodipyrrole substituted with tetraethyl is used as an illustration as follows: The starting material, 2-benzyloxycarbonyl-3,4-diethyl-5-methyl pyrrole (not shown) in acetic acid glacial is treated with lead tetraacetate. Ethylene glycol is added to reduce any remaining Pb (IV). Water and the 5-acetoxymethyl-2-benzyloxycarbonyl-3,4-diethylpyrrole shown as the starting material in the Reaction Scheme 5 are added, collected by filtration and washed with additional water. The 5-acetoxymethyl-2-benzyloxycarbonyl-3,4-diethylpyrrole is added to acetic acid in water and heated. The solid product precipitates as large pieces when the previous solution is cooled to room temperature. Water is added and the product is collected by filtration and then washed with additional water. The filtrate is extracted with CHpClp and then evaporated to produce a solid product. The solid products are combined, and then recrystallized from a solution of CH2C12 and hexanes to obtain 5, 5'-bis (benzyloxycarbonyl) -3, 3'4,4'-tetraetyl-2,2'-dipyrromethane . This product is dissolved in tetrahydrofuran (THF) and stirred under hydrogen in the presence of Pd / C and triethylamine. After the catalyst is filtered through celite, the filtrate is evaporated to dryness, dissolved in N, N-dimethylformamide and heated to boil under argon. The solution is cooled and an excess of cooled benzoyl chloride is added. The reaction mixture is stirred and the solid product is collected by filtration. The solid product is added to water and basified using NaHCO, and heated to 6 ° C. The pale yellow product, 3, 3 '4,4'-tetraetil-5,' -difor-mil-2, 2'-dipyrromethane, is crystallized from the solution and filtered and washed with water. This product dissolves in ethanol and is bubbled with argon; Hydroxylamine-hydrogen chloride and sodium acetate are added. This mixture is heated under argon and the solvent is removed and the product is dried overnight in vacuo. The resulting bisoxime is dissolved in acetic anhydride and saturated with argon to generate the crude bisnitrile product, which is obtained as a black solid after the removal of acetic anhydride and dried under vacuum. The product shown as the final product of Reaction Scheme 5 is purified by the silica gel column with 0.5% methanol in CH2C12, followed by a column of alumina with 10-20% EtOAc. Evaporation of the solvent produces 3, 3 '4,4'-tetraetyl-5, 5'-cyanodipyrrometane as pale pink crystals. The product obtained above can then be converted to the desired corresponding porpocyanine as illustrated in Reaction Scheme 2. 3,3'4,4'-Tetraethyl-5, 5'-cyanodipyrrometan in THF anyhydro is added to a THF suspension of LiAlH, under nitrogen at 0 ° C. The mixture is stirred and water is added to rapidly quench the reaction and the precipitate is filtered. The colored, golden solution is transferred to a two-necked flask containing equimolar portions of Pb (SCN) 2 and anhydrous sodium sulfate. Anhydrous methanol is added and the mixture is brought to reflux. The color changes gradually from purple to dark green. The reaction is stopped and air is slowly bubbled through the solution. The crude product is dissolved in methylene chloride and the solid is filtered. The volume of the green solution is reduced to approximately 5 ml and then loaded onto an alumina column and eluted with ethyl acetate in CHpClp. The bright green eluent containing the porfocyanine is collected and evaporated to dryness. If Reaction Scheme 3 should be followed in order to obtain the desired porfocyanine, the conduit of this sequence of reactions can be illustrated, again using the dipyrromethane substituted with tetraethyl as an example, as follows: 3, 3 '4,4'-tetraetyl-5,5'-cyanodipyrromethane in anhydrous THF is added to a THF suspension of LiAlH, under nitrogen at 0 ° C. The mixture is stirred and water is added to rapidly quench the reaction and the precipitate is filtered. 10 parts of excess 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) are added in a THF / CH 2 C 12 suspension under nitrogen at 0 ° C to 3,3 '4,4' -tetraethyl 5,5'-bisaminomethyl-2,2'-dipyrromethane in anhydrous THF / CH 2 Cl 2. The solution returns to dark green immediately. The crude product is dissolved in methylene chloride and the solid is filtered. The volume of the green solution is reduced to approximately 5 ml and then loaded onto an alumina column and eluted with ethyl acetate in CH2C12. The bright green eluant containing the porfocyanine is collected and evaporated in a vacuum. A significant increase in the yield is obtained from this synthetic process as compared to the yield obtained by means of the oxidation with air. To prepare the product of bisaldehyde and bisnitrile, 3, 3 '4,4'-tetraetyl-5,5'-cyanidipyrro methane, for example, is dissolved in THF and added to a THF suspension of LiAlH,. The resulting mixture is stirred and water is added. A solid product is formed that is filtered. The bisamine product is obtained after evaporation of the solvent by dryness under vacuum. The bisamine is dissolved in anhydrous methanol and bisaldehyde is added. The solution is bubbled and refluxed with nitrogen. Lead thiocyanate (Pb (SCNp)) is added and the solution is heated to reflux. The oxygen gas bubbles through the solution at room temperature. After evaporation of the solvent, the crude porphocyanine product is dried under vacuum. The product is purified by the column of Alp0"with ethyl acetate in CH2C12. The green eluent is collected and concentrated. The crystals of the porfocyanine product are obtained after evaporation of the solvent. The conduit is still another alternative, Reaction Scheme 4, is illustrated as follows. An illustrative dialdehyde (6) 3,3'4,4'-tetraethyl-5,5'-diformyl-2,2'-dipyrro-ethane is suspended in dry EtOH. The resulting ethanol solution is cooled to 0 ° C and ammonia gas is bubbled through it. The flask was then placed in a bath with oil at 6 ° C for 72 hours. The reaction is stopped and cooled to 0 ° C. Ethanol is removed in a rotary evaporator, the residue is chromatographed on neutral alumina with dichloromethane. This greatly simplified synthetic process produces high yields of porfocyanin. In order to prepare the alternative embodiments of the CNC expanded porforins of the invention, appropriate modifications were made to the Reaction Schemes that lead to the porfocyanine compounds per se as would be generally understood in the art. For example, to obtain the CNC extended porphyrin of the formula (1b), the condensation reactions set forth in Reaction Schemes 2 or 3 are used using 2,5-dicyanopyrrolidone as starting material. In formula (1c), the condensation of the diaminomethyl derivative with a dialdehyde is preferably used. The dialdehyde of dipyrrole methane together with the diaminomethylated pyrrolyl pyrrole are reacted to form the desired product. For the production of the compounds of the formula (1d) and (le), the CNC bonds are formed as described above from the relevant tetrapyrrole starting material. Such modi-defations can be employed which are well understood by ordinary practitioners and conventional means to synthesize these alternatives.

Administration and Use The compounds of the invention or conjugates thereof are formulated in general, in pharmaceutical compositions for administration to the subject using techniques known in the art. A summary of such pharmaceutical compositions can be found, for example, Remington's Pharmaceutical Sciences (Mark Publishing Co., Easton, Pennsylvania, latest edition). The compounds of the invention and their conjugates are administered systemically for some indications, preferably by injection. The injection can be intravenous, subcutaneous, intramuscular, or even intraperitoneal. Injectable solutions can be prepared in conventional forms, either liquid solutions or suspensions, solid form suitable for the solution or suspension in liquid before injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol and the like. Of course, these compositions may also contain minor amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffering agents and so on. Systemic administration can also be implemented through the implementation of a slow-release system or sustained release system, through suppositories, or, if formulated appropriately, orally. Formulations of these modes of administration are well known in the art, and a summary of such methods can be found, for example, in Remington's Pharmaceutical Sciences (supra). The treatment should be localized, such as for the treatment of superficial tumors or skin disorders, the compounds can be administered topically using normal topical compositions involving lotions, suspensions, pastes or creams. The amount of the compound to be administered depends on the selection of the active ingredient, the condition to be treated, the mode of administration, the individual subject, and the judgment of the practitioner. Depending on the specificity of the preparation, smaller or larger doses may be required. Doses in the range of approximately 0.05-10 mg / kg are suggested for systemic administration. Dosages in the range of approximately 0.01-20% concentration of the active ingredient, preferably 1-5%, are suggested for topical administration. A total daily dose in the range of approximately 10-300 mg is suggested for oral administration. The above ranges are only suggestive as the number of variables with respect to an individual treatment men is large and considerable deviations from these recommended values are expected. The method of image radioforming (scintigraphic image formation) of the present invention is practiced by injecting an individual parenterally with an effective cation of the porfocyanine imaging agent. By parenterally it is implied, for example, intravenously, intracavitary-intrathecally, interstitially or intracavitatively. It is contemplated that an individual will receive a dose of approximately 1 mCi at 50 mCi of the image radioforming agent, the amount is a function of the particular radioisotope and the mode of administration. For intravenous injection, the amount is usually: about 10-40 mCi, preferably about 20 mCi of Tc-99m; about 2-5 mCi, preferably about 4 mCi of In-111 or Ga-67. The image radioforming agent is conveniently provided as an injectable preparation, preferably a sterile injectable preparation for human use, to look for the agent for diseased tissue or cells, preferably comprises: a sterile injectable solution containing an effective amount of the radiolabeled agent in a sterile pharmaceutically acceptable injection vehicle, preferably phosphate buffered saline (PBS) at physiological pH and concentration. Other pharmaceutically acceptable carriers may be used as required for the site of parenteral administration. A representative preparation to be administered parenterally according to this invention, will usually contain about 0.1 to 20 mg, preferably about 20 mg, the radiolabelled agent in a sterile solution. Once sufficient isotope has been deposited at the target site, the scan is performed with either a conventional flat camera and / or SPEVT gamma, or by using a manual gamma probe used externally or internally to locate the inflation or the injury. The scintigram is usually taken by a gamma-imaging camera that has one or more windows for the detection of energies in the range of 50-500 KeV. The administration of the contrast agents for magnetic resonance imaging (MRI) is carried out in a method analogous to administration for radioforming images, except that the compounds of the invention are in the metalated forms using paramagnetic ions. Normally the signal generated correlates with the relaxation times of the magnetic moments of protons in the nuclei of the hydrogen atoms of water molecules in the region that is for imaging. The contrast agent for magnetic resonance imaging acts by increasing the rate of relaxation, thereby increasing the contrast between the water molecules in the region where the imaging agent increases and the water molecules elsewhere. in the body. However, the effect of the agent is to increase both T., and T2, the resultant trainer in higher contrast, while the latter results in the smaller constrict. Therefore, the pheomer is the concentration dependent, and there is usually an optimum concentration of a paramagnetic species for maximum efficiency. The optimal concentration will vary with the particular agent used in the image-forming site, the image-forming mode, ie echo of the spin, saturation-recovery, inversion-recovery and by various other techniques of image formation T-, dependent or Tp strongly dependent, and the composition of the medium in which the agent dissolves or is suspended. These factors, and their relative importance are known in the art. See, for example, Pykett, Scientific American (1982) 246: 78, and Runge et al., Am J Radiol (1987) 141: 1209. The MRI method of the invention is practiced by injecting an individual parenterally with an effective amount of the MRI contrast agent of the invention, which includes a metal element such as gadolinium. It is contemplated that an individual will receive a sufficient dose of contrast agent to increase the MRI signal at the target site by at least about 20%, preferably 50-500%, the amount is a function of the paramagnetic species. particular policies and the mode of administration. A contrast agent within the pre-seated invention is conveniently provided as an injectable preparation for use, preferably a sterile injectable preparation for human use, to search for an MRI agent for diseased tissues or cells, preferably comprising: a sterile injectable solution containing an effective amount of the contrast agent in a sterile pharmaceutically acceptable injection vehicle, preferably buffered phosphate buffered saline. Other pharmaceutically acceptable, conventional vehicles for parenteral administration can be used as required for the site of parenteral administration. A representative preparation to be administered parenterally according to this invention, will usually contain about 0.1 to 20 mg, preferably about 2 mg of contrast agent in a sterile solution.

And emplos The following examples are proposed to illustrate the invention but not to limit its al-canee.

Example 1 Synthesis of an Octathyl Porphocyanine, Diagram of Reaction 5 Followed by the Reduction and Formation of Imine This example describes the conduit of Reaction Scheme 5 to prepare both the bisaldehyde and dipyrrole bisnitrile intermediates. It also illustrates the condensation of bisnitrile with bisaldehyde to obtain the CNC bonds of formula (1a). Lead tetraacetate (18.2 g, 0.041 mole) is added to a stirred solution of 2-benzyloxycarbonyl-3,4-diethyl-5-methylpyrrole (10.1 g, 0.037 mole) in 60 ml of gracial acetic acid. The mixture was briefly heated to 6 ° C. Ten ml of ethylene glycol were added to reduce any remaining Pb (IC). Twenty ml of water were added and the 5-acetoxymethyl-2-benzyloxycarbonyl-3,4-diethylpyrrole (8) was collected by filtration and washed with additional water. 5-Acetoxymethyl-2-benzyloxycarbonyl-3,4-diethylpyrrole was added to 80% acetic acid in 100 ml of water and heated at 100 ° C for one hour. The solid product was precipitated as large chunks when the above solution was cooled to room temperature. One hundred ml of water was added and the product was collected by filtration and then washed with additional water. The filtrate was extracted with CHpClp and then evaporated to yield a solid product. The solid products were combined, and then recrystallized from a solution of CHpClp and hexanes to obtain 5, 5'-bis (benzyloxycarbonyl) -3,3 ', 4,4'-tetraetyl-2, 2'- dipyrromethane (7). Yield: 6.6 g, 67.4% Weight in Mol Calculated for C33H38 ° N2: 526.2831 High Resolution MS: 526.2835 1 H NMR in CDC13: 1.05 (t, 6H), 1.12 (t, 6H), 2. 43 (q, 4H), 2.75 (q, 4H), 3.85 (s, 2H), 5.25 (s, 4H), 7.28-7.40 (m, 10H), 8.70 (s broad, 2H). The above product (8.1 g, 0.015 mol) in 200 ml of tetrahydrofuran (THF) was stirred under hydrogen at atmospheric pressure and room temperature overnight in the presence of 0.4 g of 10% Pd / C and 5 drops of triethylamine. After the catalyst was filtered through celite, the filtrate was evaporated to dryness in a rotary evaporator resulting in dicarboxylic acid. The dicarboxylic acid was dissolved in 100 ml of N, N-dimethylformamide and heated to boiling under argon for one and a half hours. The solution was then cooled on ice and an excess of cooled benzoyl chloride (7.2 ml) was added dropwise and the reaction mixture was stirred for 2 hours at 5 ° C and the solid product was collected by filtration. The solid product was added to 50 ml of water and basified using NaHCO. The solution was heated and maintained at 6 ° C for one hour. The pale yellow product which was crystallized from the solution was filtered and then washed with water to obtain 3,3 '-4,4'-tetraethyl-5,5' -diformyl-2,2'-dipyrromethane (6). ) Yield: 3.4 g, 70.0% Weight in Mol Calculated for C19H26N202: 314-1994 High Resolution MS: 314.1994 1H NMR in CDC13: 1.10 (t, 6H), 1.25 (t, 6H), 2.50 (q, 4H), 2.70 (q, 4H), 4-00 (s, 2H), 9.55 (s, 2H), 10.90 (s, 2H). This product (1.03 g, 0.0033 mol) in 300 ml of ethanol was bubbled with argon for 20 minutes. Hydroxylamine-hydrogen chloride (0.51 g, 0.0073 mol) and sodium acetate (1.20 g, 0.015 mol) were added. This mixture was heated to 6 ° C under argon for two and a half hours. The solvent was removed in a rotary evaporator and the product was dried overnight in vacuo. The bisoxime was dissolved in 5 ml of acetic anhydride and saturated with argon for 30 minutes. The crude bisnitrile product (2) was obtained as a black solid after removal of acetic anhydride and dried under vacuum overnight. The product was purified by the column of silica gel (40 g) with 0.5% methanol in CHPCIpi followed by an alumina column. (40 g) with 10-20% EtOAc. Evaporation of the solvent produced pale pink crystals of 3, 3 ', 4,4'-tetraethyl-,' -cyanodipyrromethane (2). Yield: 0.43 g, 42% Weight in Mol Calculated for C19H24N4: 308.2001 High Resolution MS: 308.2002 1H NMR in CDC13: 1.10 (t, 6H), 1.25 (t, 6H), 2.45 (q, 4H), 2.60 (q , 4H), 3.90 (s, 2H), 8.45 (s, 2H). This product (2) (0.053 g, 1.7 x 10 -4"moles) was dissolved in 10 ml of anhydrous THF and added dropwise to a THF slurry of LiAlH, (0.050 g, 1.5 x 10 moles) under N2. at 0 ° C. The resulting mixture was stirred for 30 minutes and two drops of water were added in. A solid product formed, which was filtered in. The bisamine product was obtained after evaporation of the solvent by vacuum stripping overnight. The bisamine (3) was dissolved in 50 ml of anhydrous methanol and bisaldehyde (6) (0.050 g, 1.6 x 10 ~) was added.The solution was bubbled with N2 for 20 minutes and refluxed under N2. it added lead thiocyanate (Pb (SCN2)) (0.055 g, 1.7 x 10 ~ ^ moles) and the solution was heated to reflux for 4 hours.Oxygen gas was bubbled through the solution at room temperature overnight. Upon evaporation of the solvent, the crude porphocyanine product was dried under vacuum overnight. The AlpO- column was added (4% water was added) with 10% ethyl acetate in CH2C12. The green eluent was collected and concentrated in a rotary evaporator. The crystals of 2,3, 9, 10, 14,15,21, 22-octaethyl porphocyanine (1) were obtained after evaporation of the solvent. Yield: 19-3 mg, 19.1% macrocycle Weight in Calculated Mole 588.3941 High Resolution MS: 588.3933 1 H NMR (300 MHz, CDC13), -4-50 (broad s, 2H), 2.05 (t, 12H), 2.13 (t, 12H), 4-28 (q, 8H), 4-40 (q, 8H), 10.50 (s, 2H), 13.0 (s, 4H). UV / VIS in CH2C12 455, 592, 800 nm.

Example 2 Synthesis of an Octaethyl Porphocyanine. (Reaction Scheme 2) This example illustrates the formation of a porphocyanine of formula (1a) using the condensation procedure shown in Reaction Scheme 2. 3, 3 '4,4'-tetraetil-5, 5 was prepared '-cyano-pyrromethane according to the methodology of Example 1. 0.102 g, 3 > 3 x 10 - '* moles of 3, 3' 4,4'-tetraethyl-5,5'-cyananedipyrromethane in 10 ml of anhydrous THF to a suspension of 20 ml of THF of LiAlH, under N 2 at 0 ° C. The mixture was stirred for 30 minutes and two drops of water were added to rapidly quench the reaction. The precipitate was filtered. The colored, golden solution was transferred to a two-necked flask containing equimolar porcines of Pb (SCN) 2 and anhydrous sodium sulfate. Fifteen ml of anhydrous methanol were added and the mixture was brought to reflux. The color of the solution gradually changed from purple to dark green. The reaction was stopped after four and a half hours and the air bubbled slowly through the solution overnight. The crude product was dissolved in methylene chloride and the solid was filtered. The volume of the green solution was reduced to about 5 ml and then loaded onto an alumina column (4% water, 0.120 g was added) and eluted with 10% ethyl acetate in CH2C12. Two liters of the bright green eluent containing the porfocyanin were collected and evaporated to dryness. Yield: 23.4 mg, 24.1% The spectroscopic data of this compound are identical to the compound prepared in Example 1 above.

Example 3 Synthesis of an Octaethyl Porphocyanine (Reaction Scheme 3) This example illustrates Reaction Scheme 3. The 3 ', 4,4'-tetraetyl-5, 5'-cyanidipyrro methane (2) was prepared according to the methodology of Example 1. An excess of ten was added. 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) parts to 69 mg of 3,3 ', 4,4'-tetraethyl-5,5'-cyananedipyrromethane in 10 ml of anhydrous THF. The resulting solution returned to the dark green color immediately. The residue was chromatographed according to the methodology of Example 2. A significant increase in performance was obtained when compared to oxidation with air. Yield: 32 mg, 48% The spectroscopic data of this compound are identical to the compound prepared in Example 1 above.

Example 4 Synthesis of Porphocyanine (Reaction Scheme 4) This example illustrates the Reaction Scheme 4. The dialdehyde was prepared according to the method of Example 1. 25 mg of 3,3'4,4T-tetraethyl-5,5'-diformyl-2,2'-dipyrromethane (6) in 30 ml of EtOH was suspended. dry. The resulting ethanol solution was cooled to 0 ° C and the ammonia gas was bubbled through it for 30 minutes. The gas inlet was then removed and the flask was placed in an oil bath at 60 ° C for 72 hours. The reaction was stopped and cooled to 0 ° C. The ethanol was removed on the rotary evaporator and the residue was chromatographed on neutral alumina (4% HpO was added) with dichloromethane. Yield: 4.6 mg, 20% The spectroscopic data of this compound are identical to the compound prepared in Example 1 above. Another 1.5 mg of the product was obtained from the oxidation of the more polar component by DDQ.

Example 5 Synthesis of 2,2'-bis cyano meso aryl dipyrromethane This example illustrates Reaction Scheme 6. General procedure: 250 ml were loaded into a round bottom flask with methanol (115 ml), pyrrole (10 ml, 105 mmol) and p-toluenesulfonic acid (2.15 g, 11.5 mmol). A solution of the aromatic aldehyde (11.25 mmoles) in methanol (30 ml) was added to this stirred solution for 100 minutes. After this time, the reaction was poured into water (350 ml) and extracted with dichloromethane (3 x 150 ml). The organic phase was washed with brine (3 x 200 ml) and dried over anhydrous potassium carbonate. Evaporation of the solvent in vacuo followed by flash chromatography (silica gel; 100 g) of the residue of the meso aryl dipyrromethane. The mesyl aryl dipyrromethane (0.32 mmole) was dissolved in N, N-dimethylformamide (5 ml), this solution was then diluted with acetonitrile (5 ml) and the stirred mixture was cooled to -78 ° C. A solution of chlorosulfonyl isocyanate (1.42 mmol) in acetonitrile (3 ml) was added dropwise to the cooled mixture under an N2 atmosphere. The mixture was stirred at -78 ° C for 1 hour, then at -40 ° C for an additional hour before allowing it to rise to room temperature. The reaction mixture was poured into a mixture of 5% aqueous sodium hydrogen carbonate (15 ml), aqueous potassium hydroxide (3M, 2.7 ml) and ice, then diluted with brine (200 ml) and extracted with dichloromethane. (3 x 50 ml). The organic phase was dried over anhydrous potassium carbonate, the solvent was evaporated in vacuo and the residue chromatographed on silica gel to give 2,2'-bis cyano meso aryl dipyrromethane.

Example 6 Condensation of 2,2'-bis cyano meso aryl dipyrromethanes to prepare by symmetrical ocyanines This illustrates the method of Reaction Scheme 3, where at least one R. is aryl. General method: A solution of 2,2'-bis cyano meso aryl dipyrromethane (0.124 mmol) in tetrahydrofuran (5 ml) was added dropwise to a stirred suspension of lithium aluminum hydride (1.3 mmol) in tetrahydrofuran (10 ml. ) under an atmosphere of Np at 0 ° C. The reduction is followed by thin layer chromatography (silica gel), eluting with 10% ethyl acetate in dichloromethane until complete, then LiAlH, in excess, quenching with water, dichloromethane (15 ml) is added, and mixture is filtered. The filtrate is dried over anhydrous sodium sulfate and the solvent is evaporated in vacuo. The residue is dissolved again in dry dichloromethane (100 ml) and a solution of dichlorodicyanobenzoquinone (5 equivalents) in toluene (5 ml) is added dropwise to the stirred solution. Oxidation is followed by UV-vis spectroscopy and when no additional porphocyanine formation is detected, the reaction mixture is filtered through a neutral alumina plug to remove excess DDQ, and the solvent is evaporated in vacuo. The crude diphenyl porphocyanine is purified by chromatography on alumina.

Example 7 Application of the Methods of Examples 5 and 6 In the following examples which describe various porfocyanines, the following number system will be used: Using the Method of Examples 5 and 6, the following were prepared: 1. 12.24-Diphenyl porphocyanine: 1 g of benzaldehyde gives 732 mg (35%) of meso phenyl dipyrromethane after chromatography (silica gel; 10% in dichloromethane). 500 mg meso fefcyl dipyrromethane gives 195 mg (32%) of 2,2'-bis cyano meso phenyl dipyrromethane after chromatography (silica gel, 5% acetonitrile in dichloromethane). 100 mg of 2,2'-bis cyano meso phenyl dipyrromethane gives 28 mg (29%) of 1,11-diphenyl porphocyanine after chromatography (neutral alumina mesh 60-325, 10% ethyl acetate in dichloromethane). Spectroscopic data: 1H NMR (CDCl h 7.92 (m, 6H), 8.44 (m, 4H), 9-38 (d, j = 5.7 Hz, 4H), 9.8 (d, J = 5.7 Hz, 4H), 12.95 ( s, 4H); UV-vis:? 452,598,640.814nm in CH2C12; EMAR (El) for C3 / H24N6 (M +) calculated 516.2062, found 516.2058.12.24-Di (3 ', 4', 5'-trimethoxyphenyl) ) Porphocyanine: 2.21 g of 3,4-trimethoxybenzaldehyde gives 1,147 g (33%) of meso (3,4,5-trimethoxyphenyl) dipyrromethane after chromatography (silica gel, 3% acetone in dichloromethane). g of meso (3, 4, 5-trimethoxy-phenyl) dipyrromethane gives 460 mg (40%) of w, w'-bis cyano meso (3,4,5-trimethoxyphenyl) dipyrromethane after chromatography (silica gel; 10% ethyl acetate in dichloromethane) 100 mg of 2,2'-bis cyano meso (3,4,5-trimethoxyphenyl) dipyrromethane gives 31 mg (32 Z) of 1,1-di (3 ', 4' , 5'-trimethoxyphenyl) porphocyanine after chromatography (neutral alumina mesh 60-325; ethyl acetate / bichloromethane (1: 1)). Spectroscopic data: H-NMR (CDC1-) 4-1 (s, 12H), 4-25 (s, 6H), 7.67 (s, 4H), 9-44 (d, J = 4.8 Hz, 4H), 9.8 (d, J = 4-8 Hz, 4H), 12.95 (s, 4H); UV-vis:? 460,602,644,818 nm in CH2C12; HRMS (El) for C ^ H ^ N ^ (M +) calculated 696.2696, found 696.2690. 3., 24-Di (pentafluorophenyl) porfocyanine: 2.22 g of pentafluorobenzaldehyde gives 1.48 g (42%) of meso (pentafluorophenyl) dipyrromethane after chromatography (silica gel, 25% hexane in dichloromethane). 200 mg of meso (pentafluorophenyl) dipyrromethane gives 89 mg (38%) of 2,2'-bis cyano meso (penta-fluorophenyl) dipyrromethane after chromatography (silica gel, 5% ethyl acetate in dichloromethane) . 89 mg of 2,2'-bis cyano meso (pentafluorophenyl) dipyrromethanol gives 23 mg (28%) of 1,1-di (pentafluoro-phenyl) porphocyanine after chromatography (neutral alumina mesh 60-325; % in dichloromethane). 1 Spectroscopic data: H NMR ((D .-, C)? C0) $ 9. 55 (dd, J., = 15.22 Hz, J2 = 4.15 Hz, 4H), 10.1 (d, J- = 4.15 Hz, 4H), 13-24 (s, 4H); NMR 9F ((D3O2C0) 60.53, 60.92, 61.45 referenced to F3CC00H; UV-vis: 442.582.624, 814 nm in acetone, EMAR (El) for C H? / N F? O ^ M) calculated 696.1120, found 696.1124.

Example 8 Condensation to Obtain Asymmetric Porphocyanines This example uses the method of Reaction Scheme 3. General procedure: Two were dissolved 2,2'-bis cyano dipyrromethane differently substituted (0.1 mmol) in dry tetrahydrofuran (5 ml) and the two solutions were mixed. The mixed solution was added dropwise to a suspension of LiAlH, in dry THF (10 ml) under an atmosphere of 2 to 0 ° C. The reduction was followed by the CCD and, when completed, LiAlH, in excess was rapidly quenched with water and dichloromethane (5 ml) was added. The resulting slurry was filtered by gravity and the filtrate was dried over anhydrous sodium sulfate. Evaporation of the solvent in vacuo, followed by redissolution in dichloromethane (20 ml) gives a gold-colored solution. To this solution was added dropwise a solution of DDQ (5 equivalents) in toluene (5 ml). Oxidation was followed by UV-vis spectroscopy and, when no further porfocyanine formation was observed, the reaction mixture was filtered through a plug of neutral alumina. The evaporation of the solvent from the filtrate gives the crude mixture of porfocyanines, which were determined by chromatography.

Example 9 Application of the Method of Example 8 Using the method of Example 8, the following was prepared: 12-Phenyl-2,3,21,22-tetraethyl porphocyanine: 51 mg of 2,2'-bis cyano meso phenyl dipyrromethane + 58 mg of 2,2'-bis cyano 3, '4,4'-tetramethyl dipyrromethane gives 48 mg of pooled, crude porphocyanine. After chromatography (reverse phase C-.g; 20% aqueous 0.01% trifluoroacetic acid in 0.1% trifluoroacetic acid / acetonitrile) 21.7 mg (21%) of the title compound was isolated. Spectroscopic data: H NMR (CDC.) NMR 1H (CDC13) 2.05 (t, J = 7.5 Hz, 3H), 2.07 (t, J = 7.5 Hz, 3H), 4.21 (q, J = 7.5 Hz, 2H), 4-32 (q, J = 7.5 Hz , 2H), 7.88 (m, 3H), 8. 1 (m, 2H), 9-31 (d, J = 4.5 Hz, 2H), 9.7 (d, J = 4-5 Hz, 2H), 10.3 ( s, 1H), 12.72 (s, 2H), 12.96 (s, 2H); UV-vis:% 456, 592, 632, 804 nm in CH2C12; HRMS (El) for C 36 H 3 DN 6 (M) calculated 552.3001, found 552.2996.

Example 0 12.24-Diphenyl beta-alkyl porfocyanin This general procedure, which encompasses part of Reaction Scheme 5 and Reaction Scheme 2, begins with the bis carboxy dipyrromethane: 2,2'-bis carboxy 3,3'4,4'-tetraalkyl meso phenyl dipyrromethane (0.25) was dissolved mmoles) or 2,2 'bis carboxy 3, 3'-dialkyl meso phenyl dipyrromethane (0.25 mmoles) in N, N-dimethylformamide (10 ml) and the solution was heated to reflux under a stream of Ng. monitored by CCD and when decarboxylation was completed, the mixture was diluted with acetonitrile (10 ml) and cooled to -78 ° C. A solution of chlorosulfonyl isocyanate (2.9 mmol) in acetonitrile (1 ml) was added dropwise to a stirred solution under an atmosphere of N2 > The mixture was stirred for 1 hour at -78 ° C and then for an additional hour at -40 ° C before it was allowed to rise to room temperature. The reaction mixture was poured into a mixture of 5% aqueous sodium hydrogen carbonate (20 ml), aqueous potassium hydroxide (4 ml) and ice, then diluted with brine (200 ml) and extracted with dichloromethane (3 ml). x 50 ml). The organic phase was washed with brine (5 x 100 ml), dried over anhydrous potassium carbonate, and the solvent was evaporated in vacuo. Chromatography of the residue gives 2,2'-bis cyano meso phenyl-, '4,4'-tetraalkyl dipyrromethane or 2,2'-bis cyano meso phenyl-3,3'-dialkyl dipyrromethane. The bis-cyano compound (0.11 mmol) was dissolved in dry THF (10 ml) and this was added dropwise to a stirred suspension of LiAlH, (1.3 mmol) in dry THF (10 ml) under an atmosphere of 2 to 0 °. C. The reduction was monitored by the CCD and when the LiAlH / in process was completed, it was quickly cooled with water, dichloromethane (15 ml) was added and the resulting slurry was filtered by gravity. The filtrate is dried over anhydrous sodium sulfate and the solvent is evaporated in vacuo. The residue is dissolved again in dichloromethane (100 ml) and a solution of DDQ (5 equivalents) in toluene (95 ml) is added dropwise to the stirred solution. Oxidation is followed by UV-vis spectroscopy and, when no further porpocyanine formation is observed, the reaction mixture is filtered through an alumina plug. The filtrate is evaporated in vacuo and the residue is chromatographed to give 1,11-diphenyl beta-octaethyl porphocyanine or 1,11-diphenyl beta-tetraalkyl porfocyanine.

Example 11 Application of the Method of Example 10 Using the method of Example 10, the following was prepared: 12.24-Diphenyl-3,9,1, 21-tetraethyl-2,10,14,22-tetramethyl porphocyanine: 100 mg of 2,2'-bis carboxy 3, 3'-diethyl-4,4'-dimethyl meso phenyl dipyrromethane gives 40 mg (44%) of bis cyano 3, 3'-diethyl-4,4'-dimethyl meso phenyl dipyrromethane. 40 mg of bis cyano 3,3'-diethyl-4,4'-dimethyl meso phenyl dipyrromethane gives 90 μg (0.23%) of the title compound after chromatography (60-325 neutral alumina mesh; % in dichloromethane). Spectroscopic data: UV-vis: 462,604,642, 802 nm in CH2C12 EMAR (El) for C, 6H, gN6 (M +) calculated 684.3940, found 684-3932.

Example 12 Porphocyanine In Vitro Toxicity Cells are washed three times in serum free medium (DME), counted and processed at a concentration of 10 7 cells per ml. For the "affinity" test, in the dark, 100 μl of the cell suspension and 100 μl of the test or control compound are mixed. The "labeling" is allowed to continue for one hour at 4 ° C, and the labeled cells are washed in the dark three times with a medium of 3 ml each time and suspended again in a fresh medium. The resuspended cells are then subjected to exposure to light at 300-850 nm during minutes In a "direct" assay the cells are irradiated immediately upon addition of the test or control compound. The effect of irradiation is estimated using appropriate methods for fixed cells. When human erythrocytes (RBCs) are used as test cells, the hemolysis caused by the irradiation of the cells labeled with the control (hematoporphyrin, Hp) and labeled with porpocyanine (formula (1)) is visually estimated. When the murine mastocytoma cell line P815 is used, the results are determined as follows: Cells are labeled as before using concentrations of 10-50 ng / ml of Hp as control and p-phocyanine of formula (1a) as the substance test. The newly suspended cells are treated with 300-850 nm of light for 30 minutes and the resulting viability is estimated by direct counting using eosin-Y exclusion, a normal procedure to differentiate between living and dead cells. In other determinations conducted as before, cells recovered from exposure to light were tested for viability by incubating them for 18 hours in 10 μCi / ml of tritium-labeled thymidine according to the normal procedure whereby thymidine incorporation was cooled with viability. The cells are harvested and the uptake of radioactivity is measured by a scintillation counter.

Example 13 Porphocyanine Selective Link P815 cells are incubated as debrided in Example 12 using 1-200 ng / mp of Hp or the porphocyanine of formula (1a). The cells are labeled in the dark for 30 minutes, washed free of unabsorbed porphyrins, resuspended and then exposed at 300-850 nm light for another 30 minutes. The viability of the cells is established by the incorporation of tritiated thymidine after labeling with 30 μCi / ml of tritiated thymidine and incubation at 37 ° C for 18 hours.

Example 1 Preparation of Immunoconjugates This example describes methods of preparation for immunoconjugates of four different antibodies with any hematoporphyrin (Hp) or a porfocyanine (Pe) of the formula (1a). The antibodies employed are CAMAL-1, anti-M1 antibody, and B16G antibody, all prepared as described hereinabove, and the affinity of purified anti-mouse rabbit Ig (RaMIg). In addition, a resulting non-mixing monoclonal preparation is stirred for another 10 minutes. This mixture is then made by dilution in phosphate buffered saline, pH 7.4 (PBS) by adding 5 times the volume of PBS containing 50 μl of monoethanolamine, and then dialyzing against PBS using three water changes. Alternatively, 2 ml of a dipsersion containing 5 mg each of Hp or Pe, a binding agent, and a dehydrating agent are prepared and stirred for about 15 minutes at room temperature under nitrogen. To this is then added a dispersion containing about 2 mg of the immunospecific protein in 2 ml of tetrahydrofuran and the resulting mixture is stirred for another 10 minutes. The mixture is then worked up as described above. The above procedures are appropriate for CAMAL-1 and for the remaining antibody preparations listed above. In addition, the following preparations are made specifically with B16G and RrtMIg: B16G Eleven mg of hematoporphyrin plus 11 mg of EDCI are stirred in 4 ml of spectral grade DMSO during relevant, purified (C-MAb) is used where control is desired. A preparation of the conjugates is performed basically as described by Mew et al. J Immunol (1983) 130: 1473). In summary, to 220 mg of Hp. 0.2 HCl (Sigma Chemical Co., St. Louis, MO) in 25 ml of water and 0.8 ml of N, N-dimethylformamide, is added 20 ml of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide HCl (EDCI) ) in 0.6 ml of water. After 30 minutes, this solution is mixed with 15 mg of the antibody protein dissolved in 5 ml of distilled water and incubated for 5 hours. During this period, the pH of the solution is monitored and adjusted between 6 and 7. Then 50 μl of monoethanolamine is added, and the solution is allowed to stand overnight at room temperature. The solution is dialyzed against a 0.001 M phosphate buffer, pH 7.4 for four days with three changes per day and overnight against PBS. The porfocyanine conjugate is prepared analogously. In a preferred protocol, 2 ml of a dispersion in DMSO containing 5 mg each of Hp or Pe and the dehydrating agent are prepared and stirred for 30 minutes at room temperature under nitrogen. To this is added a dispersion containing 2 mg of the appropriate immunoglobulin in 2 ml of DMSO, and 30 minutes under nitrogen at room temperature before the addition of 20 mg of lyophilized B16G antibodies, prepared as described by Maier et al. J Immunol (1983) 1: 1843, in 2 ml of DMSO. The resulting mixture is stirred for 40 seconds at room temperature and worked up as described above. The resulting product contains approximately 375 μg Hp / mg B16G. A similar procedure is used that replaces Pe with Hp.

RaMIg Four hundred μg of EDCI and 400 μg of hematoporphyrin in 1 ml of DMSO are stirred for 30 minutes under nitrogen at room temperature as before, before the addition of 800 μg of lyophilized RocMIg antibodies, prepared as described by Mew et al. J. Immunol (1983) 130: 1473) in 1 ml of DMSO. The resulting mixture is stirred for 30 seconds and worked up as described above to obtain a product containing 200 gHp / mg of RMIMIg. A similar procedure was used that substitutes the Pe for Hp.

EXAMPLE 15 Specificity of Immunoconjugates In Vitrq The TSM-Hp and TSM-Pc conjugates were tested where the SST is comprised of an immunoglobulin against the cells in vitro by mixing the conjugates with the appropriate cell types, together with the appropriate controls, and then labeled cells are exposed to irradiation. The procedures for carrying out this assay are described in detail in Mew et al., Cancer Research (1985) for CAMAL-1, and by Mew et al., J. Immunol. (1983) for Anti-M1, both references cited above are incorporated herein by reference. In summary, for CAMAL-, three cell lines are marked, WC4, WC6 and WC2 (WC4 and WV6 produce the CAMAL antigen, but WC2 does not), with the appropriate TSM-Hp or TSM-Pc preparation as described in the Example 14- Preparations of labeled cells containing 10 cells each, are introduced into Rose chambers and exposed to activation with light with a laser beam at 630 nm. The results are then compiled for the various pre-parations. For the anti-Ml conjugate, Ml tumor cells are used as target cells and treated with the conjugates TSM-Hp, TSM-Pc or drug or antibody alone or the combination of antibody and drug, but without coupling, by incubating them in a incubator moistened with C02 at 6%, at 37 ° C for two hours. The cells are washed three times in PBS and then plated and exposed to fluorescent light overnight. Cells are assessed for viability by uptake of tritiated thymidine as before. For the B16C conjugates, A10, P815 and L1210 cells were used as target cells. (A10 cells are a T-cell hybridoma that secretes a supersor factor T reactive with B16G; P815 cells are also reactive with B16G.) The in vitro study is done using a direct method, using the B16G-Hp or B1óG-Pc conjugates or indirectly using unlabeled B16G antibodies and labeled R * MIg-HP or RtfMIg-Pc. - In a direct method, 5 x 10 cells are suspended in 1 ml of DME / Hepes containing the appropriate TSM-drug conjugate as a test or control in Hp or Pe concentrations of 320, 160, 80, 40 and 20 ng of drug / ml. The cells are incubated in the dark at 37CC for 1 hour, then washed 3 times in 5 ml of DME / Hepes, and then resuspended in 1 ml of the same buffer. Three test portions of 100 μl of the labeled preparations are distributed in flat bottom microtitre wells and the rest of the cell suspensions (700 μl) are exposed to 2 incandescent light (22.5 mW / cm) at a distance of 20 μl. am for 1 hour. Then three aliquots of Additional 100 μl to the microtiter wells. Then thymidine labeled with diluted tritium, in DME / Hepes containing 20% FCS, was added to all the microtiter wells in aliquots of 100 μl so that 2 μCi of labeled thymidine was added to each well. The cultures are incubated for 18 hours at 37 ° C and humidified with 10% C02 and then harvested in a MASH harvester. Thymidine incorporation is measured with a Hp scintillation counter (tri-Carb Model 4550). In an indirect assay, cells suspended in A10, prepared as described above, are exposed to 50 μg / ml of either B16G OR a control antibody C-MAb at 4 ° C for 30 minutes, washed in DME / Hepes , and then exposed for an additional 30 minutes at 4 ° C in the dark to vary the concentrations of RaMIg-Hp or R «MIg-Pc between 2 μg / ml and 15 ng / ml of Hp or Pe. The cells are then evaluated for viability using labeled thymidine uptake as described above.

Example 16 In Vivo Cytotoxicity of Porphocyanine and Conjugates of the same In vivo efficacy of porfocyanine (Pe) and conjugates thereof was also assessed. For the CAMAL-1 and anti-Ml conjugates, the procedures are described in the two sheets of Mew et al. Referenced above in Example 15. For the conjugates B16G-Hp and B16G-Pc and for the Pe (formula ( 1)) alone, the in vivo studies are conducted as follows: The in vivo test transmits the direct effect of a population of T suppressor cells on tumors, which then serve as means to assess the effectiveness of the irradiation treatment. Mastoci-P815 cells grow in syngeneic DBA / 2 mice stimulated with tumor-specific T suppressor cells. These T-suppressor cells prevent the development of specific T-killer cells that would otherwise help in the regression of the tumor. The T-cell hybridoma designated A10 above secretes a suppressor factor T which is associated with these T suppressor cells. In this way the selective killing of these populations of T suppressor cells through the reaction with conjugates in which the SST is a Specific antibody to the suppressor factor T on the surface of the cells (specifically B16G) should result in tumor regression in mice bearing the P815 tumors.

Therefore, in this assay, DBA / 2 mice are injected on the right side subcutaneously with 10 ^ of P815 cells to incorporate the tumor. On day eight, when the tumors are palpable (approximately 25-42 mm), the mice are randomly separated into groups of eight and injected IV with 150 μl of PBS containing nothing, Hp or Pe, B16G -Hp o B15G-Pc, B16G plus any drug, B16G alone or C-MAb-Hp or C-MAb-Pc. Hp levels are 50 μg per animal in all cases and B16G 310 μg in all cases (where appropriate). The animals are kept in the dark for two hours and then exposed to strong light at 300-850 nm and 22.5 mW / c. The animals are then treated normally and monitored on a daily basis.

Example 17 Diagnosis Image Formation A 32-year-old female patient develops fever and abdominal pain. The patient is maintained on antibiotic therapy for a period of one week without effect. A CAT scanner fails to demonstrate any abnormal mass. Radio-imaging studies are performed using porphocyanin labeled Tc-99m.

An injection of 20 mCi of the radiolabelled porfocyanin is used and the patient is scanned with a gamma camera in the SPECT mode. Examination of the patient's abdomen shows a focus of accumulation of Tc-99m. The surgery is performed and an abscess is found at the site of Tc-99m activity.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Having described the invention as above, property is claimed as contained in the following:

Claims (36)

1. A compound of the formula P1.Zr (Pi-Z1) B-PrZi (ll and the metallated forms and salts thereof, characterized in that it is an integer of 1-4; and wherein each P. is independently a pyrrole residue of the formula wherein each R. and R-b is independently a substituent without interference, and wherein each Z. is independently a covalent bond; or is a bridge group meso of the formula or is it a group of N-meso bridge of the formula a CC link of the formula , »R" * »< . R .. or is a CNCCNC link of the formula where each Rl.e, Ri.d-,, í4 Qe, Rí.i "and" Rí.g is indefinitely a substituent without interference, or is a CNC link of the formula where at least one Z. is the CNC link.

2. The compound according to claim 1, characterized in that n = 23.

The compound according to claim 2, characterized in that Z-, and Z are CNC links and Zp and Z, are meso bonds.

4. The compound according to claim 2, characterized in that Z * is a CNC bond and Zp, Z and Z. they are meso links.

5. The compound according to claim 2, characterized in that Z-. it's a CNC link, Zp and Z are covalent bonds, and Z is a CC bond.

6. The compound according to claim 2, characterized in that Z .. and Z ^ are CNC bonds, Zp is a colvalent bond and Z. It is a meso link.

7. The compound according to claim 2, characterized in that Z- and Z ^ are CNC bonds, Zp is a meso bond, and Z, is a CNCCNC bond.

8. The compound according to claim 2, characterized in that Z1 and Z- are CNC bonds, Zp is an N-meso bond, and Z is a meso bond.

9. The compound according to claim 2, characterized in that all Z, Z2, Z, and Z, are CNC links.

10. The compound according to claim 1, characterized in that n = 1.

11. The compound according to claim 10, characterized in that Z "is a CNC bond, and Zp and Z ,. they are meso links.

12. The compound according to claim 1, characterized in that n = 3.

13. The compound according to claim 12, characterized in that Z .. and Zp are links CNC, and Z-j, Z, and Zc are meso links.

14. The compound according to claim 1, characterized in that n = 4.

15. The compound according to claim 14, characterized in that Z1 and Z. are CNC links and Zp, Z, ¡, Z r- and Z¿ are meso bonds.

16. The compound according to claim 1, characterized in that each of R a, R-; > Rí.c, Ri.j, Rí.e, R1, -1 p and R. ?,g "is indep resistantly H, 'halo,' nitro, cyano, NR'2, SR ', OR', SOR ', SOgR1 , COOR ', C0NR'2, wherein each of R' is independently H or (1-6C) alkyl, and wherein amino, sulfhydryl and hydroxyl substituents may be optionally acylated (1-6C), optionally substituted alkyl ( 1-6C), optionally substituted (1-6C) alkenyl, optionally substituted (1-6C) alkynyl, optionally substituted aryl (4-12C), or optionally substituted arylalkyl (5-18C).

17. The compound according to claim 1, characterized in that each R. is independently optionally substituted aryl, optionally substituted aryl, optionally substituted arylalkyl or is H.

18. The compound according to claim 17, characterized in that Ric is independently optionally substituted aryl or H.

19. The compound according to claim 3, characterized in that one Rí.c is substituted aryl and the other R. is H.

20. The compound according to claim 1, characterized in that Rí.a and all Ri.b are identical.

21. The compound according to claim 3, characterized in that P-ta, Rlb) R / _ and RT are identical to each other and wherein R ~, R h, R "R are identical to each other, but different from R-1, a, Ri b, R4a and R4b '

22. The compound according to claim 21, characterized in that each of R. and R. is independently H or optionally substituted alkyl.

23. The compound according to claim 2, characterized in that it is selected from the group consisting of porfocyanine; 2.3, 9, 10, 14, 15, 21, 22-octaethylperophocyanine, 12, 24-diphenyl-phocyanine; 12,24-di- (3 ', 4', 5'-trimethoxyphenyl) porfocyanine; 12,24-di (penta-fluorophenyl) porfocyanine; 12-phenyl-2,3,21,22-tetraethyl porphocyanine, 12,24-diphenyl-2,3,9,12,12,11,22-octaethyl porphocyanine and 12,24-diphenyl-3,9, 15, 21 -tetraethyl-2, 10, 14-22-tetramethyl porpocyanine. 24- A composition of matter, characterized in that it is the compound of formula (1) linked covalently to a target-specific portion (SST).

25. The composition according to claim 24, characterized in that the covalent bond includes a linking group.

26. The composition according to claim 24, characterized in that the SST is an immunoglobulin or an immunoglobulin fragment, a steroid, or a saccharide.

27. A pharmaceutical composition, which is cytotoxic to specific cells or tissues when irradiated with light, characterized in that it comprises an effective amount of the compound of claim 1 in admixture with at least one pharmaceutically acceptable excipient.

28. A pharmaceutical composition that is cytotoxic to specific cells or tissues when irradiated with light, characterized in that it comprises an effective amount of the composition of claim 24, in admixture with at least one pharmaceutically acceptable excipient.

29. The compound according to claim 1, characterized in that it is in metalated form.

30. The compound according to claim 20, characterized in that the metal is a paramagnetic ion. • The compound according to claim 30, characterized in that the paramagnetic ion element is gadolinium (III) or manganese (II).

32. A magnetic resonance imaging (MRI) contrast agent, characterized in that it comprises the compound according to claim 1, in metalated form with a paramagnetic ion.

33. The compound according to claim 1, characterized in that it includes a radioisotope selected from the group consisting of technetium, indium and gallium.

34. A method for preparing the compound of the formula characterized in that it comprises treating a compound of the formula with a compound of the formula in the presence of DDQ in process in a non-polar aprotic solvent

35. A method for preparing the compound of the formula characterized in that the method comprises treating a compound of the formula with an excess of ammonia in a polar solvent.

36. A method for derivatizing a pyrrole nucleus with at least one cyano group, the method is characterized in that it comprises treating the pyrrole core with chlorosulfonyl isocyanate in the presence of a polar aprotic solvent. SUMMARY OF THE INVENTION Compounds of the formula are described P, -Z, - (P, -Z,) n-P, -Z, and the metallated forms and salts thereof; where n is an integer of 1-4; and wherein each P. is independently a pyrrole residue of the formula wherein R. and "R.: is independently a substituent without interference, and wherein each Z. is independently a covalent bond, or is a meso bridge group of the formula R R o L o os an N-meso bridge group of the formula a CC link of the formula R R R? E R? E or is a CNCCNC link of the formula where each R., R-jd »^ ie > ^ f ^? is independently a substituent without interference; or is a CNC link of the formula where at least one Z- is the CNC link. These compounds are useful in photodynamic therapy and diagnosis. Metalated forms are useful when the metal is paramagnetic as an MRI contrast agent.