HK1078585B - Water-soluble porphyrin platinum compounds with high tumor selectivity and their use for the treatment of benign and malignant tumor diseases - Google Patents

Description

Water-soluble porphyrin platinum compounds with high tumor selectivity and their use for treating benign and malignant tumor diseases

Technical Field

The present invention relates to novel water-soluble porphyrin platinum compounds with high tumor selectivity and their use for the treatment of benign and malignant tumor diseases. In particular, the compounds of the invention are suitable for photodynamic anti-tumour therapy in humans and mammals.

Prior Art

Complexes of platinum (II) with porphyrin ligands and their use as potent cytostatic and photodamaging antitumor agents have been described in the following publications.

W.M.Sharman, C.M.Allen and J.E.van Lier, DDT 4, (11)507-517 (1999). Photodynamic therapy: basic principles and clinical applications (Photodynamics) of the drugs.

T.okunaka and h.kato, rev.content.pharmacother, 10, 59-68 (1999). Effective application of photodynamic Therapy (photodynamic Applications of photodynamic Therapy).

H.Brunner, H.Obermeier and R.M.Szeimies, chem.Ber.1995, 128, 173-. Complexes of platinum (II) with porphyrin ligands: synergistic effects in synthesis and photodynamic therapy (platinum (II) compounds with a porphyrin ligand: synthesis and synthesis along with photodynamic therapy).

H.brunner, k. — h.schellerr and b.treittinger, inorg.chim.acta 1997, 264, 67-69. The Synthesis of hematoporphyrin-type ligands in platinum (II) complexes and in vitro assays as potent cytostatic and photodamaging antitumor agents (synthetic and viral testing of hematoporphyrin-type ligands in plastics (II) complexes).

Description of the invention

In the present invention, novel platinum porphyrin derivatives are described, which have cytotoxic properties. Surprisingly, the compounds have good water solubility and high selectivity. The compounds are useful for the treatment of cancer, in particular, for the photodynamic treatment of tumors.

The general formula of the desired compounds of the type of platinum derivatives of tetraarylporphyrins is:

formula I

X: o, S, NH, N-alkyl;

R2/R3: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r4: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r5: h, alkyl, O-alkyl, S-alkyl, halogen, nitro, cyano, amino, substituted amino;

r6: h, alkyl, O-alkyl, S-alkyl, halogen atom, nitro, cyano, amino, substituted amino.

Formula II

X: o, S, NH, N-alkyl;

R1/R2/R3/R4: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

or R2-Z-R3, wherein Z: (CH)₂)_n，n＝0-6；

R1/R4：H，-(CH₂)_n-COOR8，n＝0-6；

R5: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r6: h, alkyl, O-alkyl, S-alkyl, halogen, nitro, cyano, amino, substituted amino;

r7: h, alkyl, 0-alkyl, S-alkyl, halogen, nitro, cyano, amino, substituted amino;

r8: h, alkyl.

The general formula of the desired compounds of the hematoporphyrin platinum derivative type is:

formula III

R2/R3: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r4: h, alkyl, cycloalkyl;

r5: h, alkyl, cycloalkyl;

r6: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

structural formula IV

R1/R2/R3/R4: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

or R2-Z-R4, wherein Z: (CH)₂)_n，n＝0-6；

Or R1/R3: h, - (CH)₂)_n-COOR6，n＝0-6；

R4/R5: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r6: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl;

r7: h, alkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, cycloalkyl.

If the compounds of the invention have at least one asymmetric centre, they may be in the form: the racemates thereof, their pure enantiomers and/or their diastereomers or in the form of mixtures of these enantiomers or diastereomers.

The compounds of the invention exhibit cytotoxic activity in selected tumor cell lines. Irradiation with electromagnetic radiation having a wavelength of 600 to 730nm will enhance its anti-tumor activity. The invention therefore relates to the chemical bonding of cytotoxic substances of cis-platinum compounds with photodynamic active molecules of porphyrin derivative type in such a way that a compound of good water solubility and high selectivity is obtained.

The compounds of the invention can be administered intra-arterially, intracerebrally, intramuscularly, intraperitoneally, intrathecally, intravenously, orally, parenterally, intranasally, rectally, subcutaneously and/or topically in the form of tablets, film-coated tablets, capsules, coated tablets, powders, granules, drops, syrups, ointments, inhalation powders, infusion solutions, drinking solutions or in some other suitable form.

The medicaments contain one or several compounds in addition to conventional physiologically tolerable carriers and/or diluents or adjuvants.

The process for the preparation of medicaments is characterized in that one or several compounds are processed using customary pharmaceutical carriers and/or diluents or other auxiliaries to give pharmaceutical preparations or are brought into a therapeutically administrable form.

The synthesis of the present invention is described.

Platinum derivatives of tetraarylporphyrins

Synthesis of substituted benzaldehydes. For the reaction with 4-hydroxy-benzaldehyde, various oligo-and polyethylene glycol monomethyl ethers were activated with tosyl chloride at their alcohol ends according to literature procedures. By reacting tosylated alcohol and 4-hydroxybenzaldehyde with K in DMF₂CO₃Are refluxed together for etherification. The substituted benzaldehyde was isolated by filtration and purified by column chromatography.

With respect to the tetraarylporphyrin platinum coordination compounds to be synthesized, it is necessary to introduce two adjacent carboxylic acid groups in one of the substituted benzaldehydes. Thus, 4-hydroxybenzaldehyde is etherified with diethyl bromomalonate under alkaline conditions. Diethyl 2- (4-formylphenoxy) malonate was used with substituted benzaldehydes for the synthesis of asymmetric tetraarylporphyrins.

Synthesis of porphyrin ligand. The synthesis of asymmetric tetraarylporphyrins was performed using the Lindsey method. Pyrrole reacts with various benzaldehydes in the presence of Lewis acid catalysts to form porphyrinogen, which is oxidized to the corresponding porphyrin by p-chloranil. The tetraarylporphyrin ester was purified by several column chromatographies. By using CHCl₃And 20% methanolic KOH or the ester is hydrolyzed with pure 20% methanolic KOH solution alone to produce the carboxylic acid necessary to coordinate to the platinum (II) fragment.

Synthesis of platinum fragment. 1, 2-diaminoethane, 1, 3-diaminopropane, trans-1, 2-diaminocyclohexane and 2, 2' -bipyridine are commercially available and used as ligands for the preparation of the corresponding dichloroplatinum (II) complexes according to literature procedures. DL-2, 3-diaminopropionic acid ethyl ester dihydrochloride, L-2, 4-diaminobutyric acid ethyl ester dihydrochloride and meso-4, 5-diaminosuberic acid diethyl ester dihydrochloride were synthesized according to literature methods and used as ligands for the preparation of the corresponding diiodoplatinum (II) complexes.

And (3) synthesizing a platinum complex. For the trans-porphyrin carboxylic acidIn response, cisplatin must be activated by conversion to diammine (dihydrate) cisplatin (II) hydroxide. It is mixed with an equimolar amount of a porphyrin ligand in CHCl₃Ethanol and water or in pure water in the case of water-soluble ligands. The diammine (malonic acid) platinum (11) complex obtained precipitated. Adding CH to the reaction mixture of the water-soluble complex₂Cl₂Removing neutral impurities. The aqueous phase was evaporated to obtain the product.

The diamine (dichloro) platinum (II) fragment is activated by conversion to the diammine (dihydroxy) platinum (II) species, which reacts with an equimolar amount of each porphyrin malonic acid in CH₂Cl₂Ethanol and water or in pure water in the case of water-soluble ligands. The complex precipitates. Addition of CH to Water-soluble complexes₂Cl₂Neutral impurities are removed and the aqueous phase is then evaporated to give the product.

For the reaction with porphyrin malonic acid, the diamine (diiodo) platinum (II) complex, which is water-soluble, must be activated by conversion to the diamine (dinitrated) platinum (II) species. In this way they react with equimolar amounts of porphyrin ligand in CH₂Cl₂Ethanol and water. The water-insoluble complex precipitated after concentration of the solution.

Hematoporphyrin platinum derivative type

Synthesis of porphyrin ligand and platinum precursor. Hemin (Hemin) was transferred to protoporphyrin dimethyl ester, from which all the following reactions started. First, treatment of dimethyl protoporphyrin with 30% hydrobromic acid in acetic acid gives unstable Markownikoff adducts of HBr with two ethylene double bonds, which displace the bromine by reaction of the corresponding alkoxides with different types of alcohols. We chose hydrophilic oligo-and polyethylene glycol monomethyl ethers as alcohols. In the etherification reaction, the HBr formed catalyzes the transesterification of the methyl ester to the corresponding ester of the alcohol. The etherified hematoporphyrin ester is purified by column chromatography. The carboxylic acid required for coordination to the platinum (II) moiety was prepared by hydrolysis of the ester with 20% methanolic KOH solution.

1, 2-diaminoethane, 1, 3-diaminopropane, trans-1, 2-diaminocyclohexane and 2, 2' -bipyridine are commercially available and used as ligands for the preparation of the corresponding dichloroplatinum (II) complexes according to literature procedures. DL-2, 3-diaminopropionic acid ethyl ester dihydrochloride, L-2, 4-diaminobutyric acid ethyl ester dihydrochloride and meso-4, 5-diaminosuberic acid diethyl ester dihydrochloride were synthesized according to literature methods and used as ligands for the preparation of the corresponding diiodoplatinum (II) complexes.

And (3) synthesizing a platinum complex. The reaction of cisplatin with porphyrin carboxylic acid does not produce the desired complex. Thus, cisplatin must be activated by conversion to diammine (dihydrate) cisplatin (II) hydroxide, which reacts with an equimolar amount of porphyrin ligand in a mixture of ethanol and water, or in the case of water-soluble ligands in pure water. The platinum (11) diammine (dicarboxylic acid) complex obtained precipitates. Adding CH to the reaction mixture of the water-soluble complex₂Cl₂Neutral impurities are removed and the aqueous phase is then evaporated to give the product.

The diamine (dichloro) platinum (II) precursor is activated by conversion to the diammine (dihydroxy) platinum (II) species, which is reacted with an equimolar amount of the respective porphyrin carboxylic acid in a mixture of ethanol and water, or in the case of water-soluble ligands in pure water₂Cl₂Neutral impurities were removed and the aqueous phase was evaporated to yield the product.

For reaction with porphyrin carboxylic acids, the diamine (diiodo) platinum (II) complex, which is water soluble, must be activated by conversion to the diamine (dinitrated) platinum (II) species. They are reacted in this way with equimolar amounts of porphyrin ligand in a mixture of ethanol and water or, in the case of water-soluble ligands, in pure water. The water-insoluble complex precipitated after concentration of the solution. The water soluble complex was isolated by chromatography on silica.

Illustrative embodiments

The following examples are intended to illustrate the invention in more detail.

The compounds of the present invention are the tetraarylporphyrin platinum derivatives exemplified in examples 1 and 2, and the hematoporphyrin platinum derivatives exemplified in examples 3, 4 and 5.

Example (b):

example 1

Diammine [2- (4- {10, 15, 20-tris [4- (1, 4, 7-trioxabinyl) phenyl ] porphyrin-5-yl } phenoxy) malonic acid (malonato) ] platinum (II) (No. 21 in FIG. 1)

The compound 2- (4- {10, 15, 20-tri [4- (1, 4, 7-trioxa octyl) phenyl]Porphyrin-5-yl } phenoxy) malonic acid (109mg, 0.100 mmol) dissolved in 10 ml of CHCl₃And 20 ml of EtOH, mixed with 0.100 mmol of aqueous diammine (dihydrate) cisplatin (II) hydroxide solution and stirred for 20 hours. Yield: 81.0 mg (54.2. mu. mol, 54%) of a violet powder, mp 213-.

Elemental analysis value (C)₆₂H₆₆N₆O₁₄Pt·10H₂O, 1494, 5) C: calculated values 49, 83; found 49, 19.H, N: calculated values: 5, 62; found 6.09.

Example 2

(±) -trans-1, 2-diaminocyclohexane [2- (4- {10, 15, 20-tris [4- (1, 4, 7, 10-tetraoxaundecyl) phenyl ] porphyrin-5-yl } phenoxy) malonic acid ] platinum (II) (No. 29 in fig. 1).

Make 10 ml of CH₂Cl₂And 122 mg (0.100 mmol) of the compound 2- (4- {10, 15, 20-tris [4- (1, 4, 7, 10-tetraoxaundecyl) phenyl in 20 ml of EtOH]Porphyrin-5-yl } phenoxy) propanThe diacid was reacted with 0.100 mmole of activated (±) trans-1, 2-diaminocyclohexane (dichloro) platinum (II). Yield: 113 mg (73.9. mu. mol, 74%) of a violet solid, mp 208 ℃. Elemental analysis value (C)₇₄H₈₆N₆O₁₇Pt，1526.6)C，H，N。

Example 3

Diammine {7, 12-bis [1- (1, 4, 7-trioxoctyl) ethyl ] -3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid } platinum (II) (No. 21 in FIG. 2).

The compound 7, 12-bis [1- (1, 4, 7-trioxa octyl) ethyl]-3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid (80.3 mg, 0.100 mmol) was dissolved in 6 ml of EtOH, mixed with 0.100 mmol of diammine (dihydrate) cisplatin (II) hydroxide aqueous solution and stirred for 20 hours. Yield: 23.0 mg (22.3. mu. mol, 22%) of a dark brown powder, mp > 250 ℃. Elemental analysis value (C)₄₄H₆₂N₆O₁₀Pt, 1030.1). C: calculated value 51.30; found 50.75. H: calculated value 6.07; found value of 5.49.N

Example 4

(±) -trans-1, 2-diaminocyclohexane {7, 12-bis [1- (1, 4, 7-10, 13, 16-hexaoxaheptadecyl) ethyl ] -3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid } platinum (II) (No. 38 in fig. 2).

The compound 7, 12-bis [1- (1, 4, 7-10, 13, 16-hexaoxaheptadecyl) ethyl ] -3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid (107 mg, 0.100 mmol) was dissolved in 10 ml of EtOH and reacted with 0.100 mmol of activated (±) -trans-1, 2-diaminocyclohexane (dichloro) platinum (II).

Yield: 25.5 mg (17.2. mu. mol, 17%) of a reddish brown powder; mp 245 ℃: elemental analysis value (C)₆₂H₉₄N₆O₁₆Pt·6H₂O, 1482.6) C: calculated 50.23; found 49.02. H: calculated 7.21; found 6.33. N: calculated values 5, 67; found 6.41.

Example 5

2, 2' -bipyridine {7, 12-bis [1- (1, 4, 7-trioxoctyl) ethyl ] -3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid } platinum (II) (No. 40a in FIG. 2).

42.2 mg (0.100 mmol) of the compound 2, 2' -bipyridine (dichloro) platinum (II) are suspended in 15 ml of H₂And (4) in O. After 10 minutes sonication was added 34.0 mg (0.200 mmol) of AgNO₃And the reaction mixture was stirred at room temperature for 4 hours under protection from light. The precipitated AgCl was filtered and washed with water. The filtrate containing the activated platinum (II) complex is evaporated. The residue was dissolved in 5 ml of H₂O and with 80.3 mg (0.100 mmol) of 7, 12-bis [1- (1, 4, 7-trioxoctyl) ethyl in 10 ml of EtOH]-3, 8, 13, 17-tetramethylporphyrin-2, 18-dipropionic acid. After stirring for 20 hours at 50 ℃ and cooling to room temperature, the precipitated solid was filtered, washed with water and EtOH and dried in vacuo.

Yield: 64.0 mg (55.5. mu. mol, 55%) of a dark pink powder, mp > 250 ℃. Analytical value (C)₅₄H₆₄N₆O₁₀Pt，1152.2)C，H，N。

Biometric data

For example, cytotoxicity effect data were obtained with human tumor cell lines TCC-SUP and J82. The effect of the compounds was investigated under protection from light and under illumination at a wavelength of 600-730 nm. It goes without saying that the compounds selected under light are more cytotoxic. There is synergy between the cytotoxic effects of the platinum component and the photodynamic principle.

Cell lines and general methods

To determine the antiproliferative activity of the novel porphyrin ligands and the corresponding platinum complexes with two different amine non-leaving groups, two bladder cancer cell lines TCC-SUP and J82 were selected as in vitro models.

To distinguish between cytotoxic and phototoxic effects, all experiments were performed twice. Cells were seeded onto microplates and test compounds were added after 48 hours. A batch of microplates was kept dark until the end of the experiment, at the same time, at 24J-cm 48 hours after addition of material^-2The other microplate was irradiated with the light dose for 10 minutes, and then the plate was re-incubated under dark conditions.

End-point chemical sensitivity analysis

Hematoporphyrin platinum derivative type

At a dose of 1 μ M, the type of non-leaving group affected the photophobic-and phototoxicity of the porphyrin-platinum conjugate. After protection from light and irradiation, the platinum complexes with 2, 2' -bipyridine (40, 41), ethyl DL-2, 3-diaminopropionate (42-46), ethyl DL-2, 3-diaminobutyrate (47-51), diethyl meso-4, 5-diaminosuberate (52-55) ligands at a concentration of 1. mu.M were inactive. Compounds with 1, 2-diaminoethane (27-30) and 1, 2-diaminopropane (31-34) leaving groups are also inactive against TCC-SUP cells. The porphyrin-platinum conjugates of most interest are those with diammine (21-26) and (. + -.) -trans-1, 2-diaminocyclohexane (35-39) ligands. Of these series of compounds, the water-soluble complexes 26 and 39 are the most active, T/C_corr.About 30% and 15%, respectively, and the reference cisplatin had a T/C of about 2% at a concentration of 1. mu.M_corr.The value is obtained. At this dose, there was no statistically significant increase in cytotoxicity of the irradiation of bladder cancer cells.

Increasing the concentration of complexes 40-55 to 5 μ M did not result in an increase or only minimally increased photophobic toxicity (FIG. 2). Most of these complexes were not much more phototoxic than the cytotoxicity observed without irradiation. However, there were significant effects for 42, 45, 47, 49, 50 and 53, while for 40 and 44, a very strong effect on TCC-SUP cell proliferation under irradiation was observed (figure 2). For compound 52, the greatest synergistic effect was found to result in tumor cell lysis.

In addition to cisplatin, the highest antitumor activity was determined for the series of porphyrin-platinum conjugates with non-leaving groups of diammine (21-26) and (geo) -trans-1, 2-diaminocyclohexane (35-39). The difference between photophobia and light-induced toxicity is best for water-soluble porphyrin-platinum conjugates 26 and 39 with n ≈ 17 side chain lengths at positions 7 and 12 of the porphyrin leaving group. All of the ethylenediamine and propylenediamine complexes 27-34 showed significant light-induced toxicity (fig. 2).

Platinum derivatives of tetraarylporphyrins

At doses of 1 μ M and 5 μ M, the type of non-leaving group highly affected the photophobic-and phototoxicity of the tetraarylporphyrin-platinum conjugates 21-38, and the results were identical to those of the hematoporphyrin-platinum complexes discussed above. 23, 29 and 30 are the most active tetraarylporphyrin-platinum conjugates, at a concentration of 1. mu.M, T/C_corr.The values were about 37%, 57% and 63%, respectively. This is similar to the hematoporphyrin-platinum complex, the most active being those with a non-leaving group of diammine or (±) -trans-1, 2-diaminocyclohexane. There was only a slight increase in cytotoxicity of the tetraarylporphyrin-platinum conjugates with side chain lengths of n-2 and n-3 under irradiation at a concentration of 1 μ M. On average, light-induced T/C_corr.The values were approximately 20% less toxic than photophobic cells (data not shown).

Increasing the complex concentration to 5 μ M enhanced the photophobic effect and phototoxicity as shown in figure 1. In addition to cisplatin, the highest antitumor activity was measured for the tetraarylporphyrin-platinum conjugate with non-leaving groups of diammine (21-23) and (+ -) -trans-1, 2-diaminocyclohexane (28-30). The difference between photophobic and light-induced toxicity was best for tetraarylporphyrin-platinum complexes 24, 27, 32-34, 36 and 38 with n-2 and n-3 side chain lengths (fig. 1).

Claims (6)

1. The following porphyrin platinum derivatives:

a platinum derivative of a tetraarylporphyrin according to formula I:

n: 2, 3 or 17

X is O;

r2 and R3 are H;

r4 is-CH₃；

R5 is H;

r6 is H;

or a platinum derivative of tetraarylporphyrin according to formula II:

n: 2, 3 or 17

X is O;

r7 and R10 are H,

r8 and R9 are- (CH)₂) And R8 and R9 together with Z form a ring, wherein Z ═ CH (CH)₂)₂；

Alternatively, the first and second electrodes may be,

r7 and R10 are each independently H, -COOC₂H₅Or- (CH)₂)₂-COOC₂H₅，

R8 and R9 are H,

z is absent;

r11 is-CH₃；

R12 and R13 are H.

2. The use of a compound according to claim 1 as therapeutically active compound for the preparation of a medicament for the treatment of bladder cancer.

3. The use according to claim 2, wherein the medicament is a medicament for photodynamic therapy of bladder cancer.

4. Use according to claim 2 or 3, wherein the medicament is a medicament for photodynamic therapy of bladder cancer by irradiation with electromagnetic radiation having a wavelength of 600 to 730 nm.

5. Medicament containing one or several compounds according to claim 1 and conventional physiologically tolerable carriers and/or diluents or adjuvants.

6. Process for the preparation of a medicament according to claim 5, characterized in that one or several compounds according to claim 1 are processed using customary pharmaceutical carriers and/or diluents or other auxiliaries to give pharmaceutical preparations or are brought into a therapeutically administrable form.