HU208486B - Process for producing pharmaceutical compositions containing pteridine derivatives - Google Patents

Abstract

The present invention relates to the preparation of a pharmaceutical composition comprising a pteridine derivative of the formula I as an active ingredient and suitable for the treatment of tumor resistance to chemotherapeutic agents. Wherein R 3 is a straight or branched C 2-6 alkyl group, optionally substituted with a carbon atom adjacent to the nitrogen atom, substituted with a hydroxyl group, R 4 is a C 1 -C 4 alkyl group, wherein one carbon atom is adjacent to the carbon atom adjacent to the nitrogen atom; R2 and R7 are morpholino or piperidino substituted with one or two methyl or ethyl groups. EN 208 486 B Scope of the description: 8 pages (including 7 sheets)

Description

FIELD OF THE INVENTION The present invention relates to pharmaceutical compositions containing pteridine derivatives of formula (I), their geometric or optical isomers, and their physiologically tolerable acid addition salts, which are effective against resistance to natural chemotherapeutic agents.

In the general formula (I)

R _{3 is a} straight or branched C 2 -C 6 alkyl group optionally substituted by one carbon atom other than the carbon atom adjacent to the nitrogen atom,

R ₄ is a C 1-4 alkyl group in which one carbon atom other than the nitrogen atom is substituted with a hydroxy group,

R ₂ and R ₇ represents a morpholino or piperidino substituted with one or two methyl or ethyl groups.

DE-A-3 445 298 discloses 2-piperazinopteridines which, inter alia, have inhibitory effects on tumor metastases and tumor development.

For the treatment of neoplastic diseases, there are, inter alia, medicinal products of natural origin such as vinca alkaloids such as vinblastine, vincristine or vindesine, epipodophyllotoxins such as etoposide or teniposide, and antibiotics such as Pharmacological Basis of Therapeutics, Macmillan Publishing Company, New York,

7th Edition, 1240-1247. and 1277-1289. (1985)]. During chemotherapeutic treatments with these agents, it has often been found that the tumors to be treated do not respond to treatment due to primary resistance or secondary resistance developed during previous treatment, or that treatment-resistant tumor cells may remain latent after tumor remission. These resistant tumor cells generally subsequently trigger metastases.

It has been found that pharmaceutical compositions containing the above-described pteridine derivatives of the formula I, their geometric or optical isomers, and their physiologically tolerated acid addition salts, induce remission of resistant tumors.

The tumor cells may be sensitized to resistance to chemotherapeutic agents by administration of the compounds of formula (I) or their physiologically tolerated acid addition salts prior to, concurrently or subsequently with the administration of said chemotherapeutic agents.

The present invention therefore relates to pharmaceutical compositions comprising as active ingredient a pteridine derivative or a geometric or optical isomer thereof and a physiologically acceptable acid addition salt thereof, optionally together with a chemotherapeutic agent or a chemotherapeutic agent of natural origin. processed to a secondary tumor resistance drug.

Thus, the pharmaceutical compositions of the present invention sensitize resistant tumor cells to chemotherapeutic agents.

The compounds of formula (I) and their preparation are described in part in U.S. Patent Nos. CA-A-912,556, US-A-3,557,105 and US-A-3,574,206, and in part in U.S. Patent No. HU-B205,761. . The known compounds have antithrombotic activity.

The compounds of formula (I) may be prepared by the methods described in the above-mentioned publications. For example, in one method, a pteridine of formula II, wherein R ₃ and R ₄ and n are defined above, one of Z ₂ and Z ₇ being a nucleophilic exchangeable group and the other of R ₂ and R _{7 being as} defined above, The compound of formula (III) wherein X is R ₂ or R _{7 is as} defined above, and if desired, separating the resulting compound into its optical or geometric isomers or converting it into a physiologically acceptable acid addition salt of an inorganic or organic acid.

Examples of substituents in the compound of formula (I) include:

R ₂ and R ₇ are independently selected from 2-methyl-piperidino, 3-methyl-piperidino, 4-methylpiperidino, 2-ethylpiperidino, 3-ethylpiperidino, 4-ethylpiperidino, 3,5 -dimethyl-piperidino, 3,5-diethyl-piperidino, 2-methylmorpholino, 2-ethylmorpholino, 3-methylmorpholine,

3-ethylmorpholino, 2,6-dimethylmorpholino, cis-2,6-diethylmorpholino, cis-2,6-diethylmorpholino, trans2,6-diethylmorpholino, 3,5-dimethyl you are morpholino

3,5-diethyl-morpholino;

R _{3 is} 2-hydroxyethyl, 2-hydroxy-n-propyl, 2-hydroxyisopropyl, 3-hydroxypropyl, 4-hydroxy-n-butyl, 2-hydroxy-2-methyl-n -propyl, 5-hydroxy-n-pentyl, 6-hydroxy-n-hexyl, 2,3-dihydroxy-n-propyl, methyl, ethyl, npropyl or isopropyl,

R ₄ 2-hydroxyethyl, 2-hydroxy-n-propyl, 3-hydroxypropyl, 4-hydroxy-n-butyl or 2-hydroxy-2-methyl-n-propyl.

Preferred are the pteridines of formula I wherein

R ₂ and R ₇ are independently 3,5-dimethylpiperidino, 2-methylmorpholine, 2,6-dimethylmorpholino, cis-2,6-dimethylmorpholino or trans-2,6-dimethyl -morfolinocsoport,

R _{3 is} methyl, ethyl, 2-hydroxyethyl, 2-hydroxy-n-propyl, 2,3-dihydroxy-n-propyl or 2-methyl-2-hydroxypropyl;

R ₄ 2-hydroxyethyl, 2-hydroxy-n-propyl or 2-methyl-2-n-propyl group and the optical and geometric isomers thereof, and physiologically tolerated acid addition salts thereof.

The following compounds have been found to be particularly effective:

4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] 2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine,

4- (N-ethyl-amino ethanol) -2,7-bis (2,6-dimethyl-morpholino) -6-phenyl-pteridine,

HU 208 486 Β

4- [ethanol-isopropanol-amino) -2,7-bis (2,6-dimethylmorpholino) -6-phenyl-pteridine,

4- [bis (2-hydroxy-2-methyl-n-propyl) amino] -2,7bisz (2,6-dimethyl-morpholino) -6-phenyl-pteridine,

4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] 2,7-bis (3,5-dimethylpiperidino) -6-phenylpteridine,

4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] 2,7-bis (cis-2,6-dimethylmorpholino) -6-phenylpteridine,

4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] 2,7-bis (trans-2,6-dimethylmorpholino) -6-phenylpteridine,

4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2- (2,6-dimethylmorpholino) -7-morpholino-6-phenylpteridine and

4- [N- (2-Hydroxy-2-methyl-n-propyl) ethanolamino] -7- (2,6-dimethylmorpholino) -2-morpholino-6-phenylpteridine and their physiologically acceptable acid addition salts .

As an example,

A = 4- [N- (2-hydroxy-2-methylpropyl) ethanol amino] -2,7bisz (2,6-dimethyl-morpholino) -6-phenyl-pteridine,

B = 4- (N-ethyl-isopropanol-amino) -2,7-bis (2,6-dimethylmorpholino) -6-phenyl-pteridine,

D = 4- (diethanolamino) -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine and

E = 4- [N- (2-hydroxy-2-methylpropyl) ethanolamino] -2,7bis (cis-2,6-dimethylmorpholino) -6-phenylpteridine,

The sensitizing effect of F = 4- [N- (2-hydroxy-2-methylpropyl) -ethanolamino] -2,7bis (3,5-dimethylpiperidino) -6-phenylpteridine compounds, e.g., adriamycin-resistant cells, as follows:

Adherent adriamycin-resistant S 180 murine sarcoma cells were cultured for 6 days in the presence of various concentrations of test substances. Cytotoxic or cytostatic concentrations of test substances are determined based on reduced cell growth or cell death. At the end of the assay, the number of live cells in each culture was indirectly measured using the property of living cells that MTT [= 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] they are capable of reducing dyestuffs to colored formazan. The concentration of test substance is considered to be the IC ₅₀ value which can reduce the number of vital cells in each culture to half of the untreated controls. The effects of the substances were also investigated in the absence of adriamycin and in the presence of non-inhibiting amounts of adriamycin under culture conditions. This gives 2 IC ₅₀ values for each test substance, one in the presence of Adriamycin (IC ₅₀ ADR) and the other in the absence of Adriamycin (ID ₅₀ ). The difference between the two IC _50s is the logarithm of the tens based A = IgIC50 to IgIC50 ADR is the measure of the increase in cytotoxicity of the test substance by adriamycin.

Description of the experiment:

2000 to 2000 exponentially growing, 100 µl medium (RPMI-1640 containing 10% fetal bovine serum) containing adriamycin-resistant or susceptible S 180 cells was plated in 96-well inoculated wells. Incubated at 37 ^° C, 5% carbon dioxide and 100% humidity Atenyészlemezt oven. After 24 hours, 5050 µl of medium containing various concentrations of the test substance and 50 µl of Adriamycin containing media are added to the cultures. After an additional 6 days of culture, 50 µl of saline solution containing 5 µl of tetrazolium [5 mg 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] ml of phosphate buffer before use in RPMI Diluted with -1640) and pipetted into each well. The plates are shaken briefly and the optical thickness measured at 570 nm using, for example, a Dynatech MR-600 type photometer. After 4 hours of incubation, the culture medium is carefully aspirated and the intracellular formazan is dissolved in 150 µl / well of dimethyl sulfoxide. The amount of colored formazan formed by reduction of the tetrazolium salt is proportional to the number of living cells. The mean of three determinations is used to calculate the IC ₅₀ (dilution 1: 2).

Table 1

Material IC ₅₀ [pg / ml] Adriamycin (ng / ml) lg ^{an IC} ₅₀ IC 25 ng Adriamycin 0 25 THE 20.69 1.49 1.14 B 31.60 4.43 0.85 C 12.85 3.77 0.53 D 14.37 5.78 0.40 E 7.50 0.50 1.18

Table 2

Material IC ₅₀ [µg / ml] Adriamycin (ng / ml) ic ₅₀ lg IC ₅₀ 1 00 ng adriamycin 0 100 THE 20.69 0.50 1.61 B 31.60 0.77 1.61, C 12.85 0.82 1.20 D 14.37 1.57 0.96 E 7.50 0.10 1.88 F 1.18

The pteridines of formula (I) above, as shown, expressly sensitize adriamycin-resistant sarcoma cells, and are thus useful in combining antimicrobial therapy with vinca alkaloids, epipodophyllotoxins or antibiotics such as daunorubicin, doxorubicin, they eliminate their resistance and thus result in the remission of tumors resistant to these substances. Tumo3 is sensitive to said chemotherapeutic agents

In the case of HU 208,486 p r, the pteridines of the formula I, together with the chemotherapeutic agents, also prevent the treatment of the resistant tumor cell populations and relapse.

Apteridines, which are otherwise well tolerated, may be administered alone or in combination with a conventional dose of chemotherapeutic agent; the dose of pteridine is 1-50 mg / kg body weight / day, preferably 320 mg / kg body weight / day, divided into 1-4 unit doses.

When used in combination with a suitable chemotherapeutic eye, a simultaneous administration of an intravenous dosage form, such as an ampoule, and tablets, dragees, suspensions, solutions, capsules or suppositories, may be contemplated.

Natural substances introduced into the chemotherapy of neoplastic diseases [cf. Goodman and Gilman, The Pharmacological Basis of Therapeutics, Macimban Publishing Company, New York, 7th Edition, 1240-1247, and 1277-1289. (1985)], the first use of a pteridine or a physiologically acceptable acid addition salt thereof of formula I is preferably carried out with or prior to the chemotherapeutic agent used or containing at least one of the aforementioned chemotherapeutic agents (see DeVita et al. in the monograph Cancer, Principles and Practice of Oncology, 2nd Edition, JB Lippincott Company Philadelphia) with or without a combination of multiple chemotherapeutic agents.

Otherwise, a compound of formula (I) or a physiologically tolerable salt thereof may be administered orally or intravenously, as appropriate.

A suitable combination for intravenous administration according to the invention preferably comprises from 1 to 25 mg / kg, preferably from 1 to 20 mg / kg body weight of pteridine or a physiologically tolerated acid addition salt thereof and a suitable chemotherapeutic agent or a combination of various suitable chemotherapeutic agents. For example, vinblastine 0.1-0.15 mg / kg every 7 days, when the dose may be gradually increased by 0.05 mg / kg, depending on the degree of side effects, is approx. 2 mg / m ² body surface area of vincristine for each 7 days for juvenile leukemia, while in adults, therapy is preferably initiated at a dose of 0.01 mg / kg every 7 days and 0.02-0.05 mg / day depending on tolerability. can be increased up to kg,

3-4 mg / m ² body surface area of vindezine every 7 days,

25-30 pg / kg mitramycin daily or for 2 days

60-75 mg / m ² body surface area of Adriamycin every 21 days,

30-60 mg / m ² body surface area daunorubicin daily for 3 days,

10 to 15 µg / kg dactinomycin daily for 5 days,

50-100 mg / m ² body surface area etoposide daily for 5 days and mg / m ² body surface area teniposide daily for 5 days, with a 10-day break always followed by 6-10 treatments.

The invention is further illustrated by the following examples.

/. Example 4: Pills containing 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino) -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

agent 75.0 mg Calcium phosphate 93.0 mg Corn starch 35.5 mg polyvinylpyrrolidone 10.0 mg Hydroxypropyl methylcellulose 15.0 mg Magnesium stearate 1.5 mg

230.0 mg

manufacture:

The active ingredient is mixed with calcium phosphate, corn starch, polyvinylpyrrolidone, hydroxypropylmethylcellulose and half of the stated amount of magnesium stearate. On a tableting machine kg. It is pressed into particles having a diameter of 13 mm, passed through a sieve with a mesh size of 1.5 mm using a suitable machine and then mixed with the remaining magnesium stearate. This granulate is compressed into tablets of the desired shape on a tabletting machine.

Seed weight: 230 mg

Print head: 9 mm, convex.

The dragee cores thus obtained are coated with a film consisting essentially of hydroxypropyl methylcellulose. The finished film coated dragee is polished with beeswax.

Powder weight: 245 mg.

II. example

100 mg 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

Composition of one tablet:

Active ingredient 100.0 mg

Milk sugar 80.0 mg

Corn starch 34.0 mg

Polyvinylpyrrolidone 4.0 mg

Magnesium stearate 2.0 mg

220.0 mg

manufacture:

The active ingredient, lactose and starch are uniformly moistened with an aqueous solution of polyvinylpyrrolidone. The wet mass is passed through a 2.0 mm sieve and, after drying in a tray oven at 50 ° C, is again sieved through a 1.5 mm sieve and mixed with the lubricant. The compression mixture is processed into tablets.

Tablet weight: 220 mg

Diameter: 10 mm, biplane with bevel on both sides and divider on one side.

Example 111

Tablets containing 150 mg of 4- [N- (2-Hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

Composition of one tablet:

Active ingredient 150.0 mg

Milk sugar powder 89.0 mg

Corn starch 40.0 mg

Colloidal Hard Acid 10.0 mg

HU 208 486 Β

Polyvinylpyrrolidone 10.0 mg

Magnesium stearate 1.0 mg

300.0 mg

manufacture:

The active ingredient, mixed with lactose, corn starch and silica, is moistened with a 20% aqueous solution of polyvinylpyrrolidone and passed through a sieve with a mesh of 1.5 mm.

The granules dried at 45 ° C are sieved once more and mixed with a certain amount of magnesium stearate. The mixture is compressed into tablets.

Tablet weight: 300 mg

Print head: 10 mm, flat

ARC. example

Hard gelatin capsules containing 150 mg of 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino-2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

Capsule composition:

Active ingredient 150.0 mg

Corn starch, dried approx. 180.0 mg Lactose powder, 87.0 mg

Magnesium stearate 3.0 mg

420.0 mg

manufacture:

The active ingredient is mixed with the excipients, passed through a 0.75 mm sieve and homogeneously mixed in a suitable apparatus.

The resulting mixture is filled into size 1 hard gelatin capsules.

Capsule Capacity: Approx. 320 mg

Capsule Shell: Size 1 hard gelatin capsule

Example V

Suppositories containing 150 mg of 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine suppository:

Active ingredient 150.0 mg

Polyethylene glycol 1500 550.0 mg

Polyethylene glycol 6000 460.0 mg

Polyoxyethylene sorbitan monostearate 840.0 mg

2,000.0 mg

manufacture:

After melting the suppository, the active ingredient is uniformly distributed therein and the melt is poured into pre-cooled molds.

VI. EXAMPLE 1 mg suspension of 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino- (2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine)

100 ml of suspension:

agent 1.0 g Carboxymethyl cellulose sodium salt 0.1 g p-Hydroxy-benzoic acid methyl ester 0.05 g p-hydroxybenzoic acid propyl ester 0.01 g Cane sugar 5.0 g Glycerol 5.0 g

Sorbitol solution, 70% 20.0 g

Aroma 0.3 g

Make up to 100 ml with distilled water

manufacture:

Distilled water is heated to 70 ° C. While stirring, the methyl p-hydroxybenzoic acid methyl ester and propyl ester as well as the glycerol and the carboxymethyl cellulose sodium salt were dissolved therein. The mixture is cooled to room temperature and the active ingredient is added with stirring and then homogeneously dispersed. After adding and dissolving sugars, sorbitol solution and flavor, the suspension is evacuated with stirring to remove air.

Each ml of suspension contains 50 mg of the active substance.

VII. Example Ampoules containing 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (cis-2,6-dimethylmorpholino) -6-phenylpteridine

Ingredients:

Active ingredient 10.0 mg

0.01 N hydrochloric acid s. q.

Make up to 2.0 ml with double-distilled water.

manufacture:

Dissolve the active ingredient in a sufficient amount of 0.01 N hydrochloric acid, make the solution isotonic with saline, filter sterile and fill into 2 ml ampoules. Sterilization was carried out by heating at 121 ° C for 20 minutes.

VIII. Example Ampoules containing 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

Ingredients:

Active ingredient 50.0 mg

0.01 N hydrochloric acid s. q.

Make up to 10.0 ml with double-distilled water.

manufacture:

Dissolve the active ingredient in a sufficient amount of 0.01 N hydrochloric acid, make the solution isotonic with saline, filter sterile and fill into 10 ml ampoules. Sterilization was carried out by heating at 121 ° C for 20 minutes.

IX. EXAMPLE EXAMPLE Dry Ampoules containing mg doxorubicin and 10 mg 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine

Composition of dry ampoules:

Doxorubicin 10.0 mg

Active ingredient 10.0 mg

Preparation

Both drugs are dissolved in the required amount of 0.01 N hydrochloric acid, sterile filtered and lyophilized.

Example X Dry powder containing doxorubicin mg and 50 mg 4- [N- (2-hydroxy-2-methyl-n-propyl) ethanolamino] -2,7-bis (2,6-dimethylmorpholino) -6-phenylpteridine vials

HU 208 486 Β

Composition of dry ampoules:

Doxorubicin 50.0 mg

Active ingredient 50.0 mg

manufacture:

Both materials are dissolved in the required amount of 0.01 N hydrochloric acid, sterile filtered and lyophilized.

The solvent ampoule contains 25 ml of saline.

Before use, the lyophilisate is dissolved in sterile physiological saline.

Of course, all other compounds of formula I can be used as active ingredients in the galenic formulations described.

Claims (4)

PATENT CLAIMS CLAIMS 1. A process for the preparation of a pteridine derivative of the Formula I wherein R _{3 is a} straight or branched C 2 -C 6 alkyl group optionally substituted with one carbon atom of a hydroxy group other than a carbon atom adjacent to the nitrogen atom. R ₄ is a C 1 -C 4 alkyl group in which one carbon atom other than the nitrogen atom is substituted by a hydroxy group. R ₂ and R ₇ for the preparation of pharmaceutical compositions containing one or two methyl or ethyl substituted morpholino or piperidino groups, an optical or geometric isomer thereof, and a physiologically acceptable acid addition salt thereof, characterized in that the active ingredient is prepared in combination with a chemotherapeutic agent of natural origin or, when R ₂ and R _{7 are the} same or substituted in the 2-position or 2,6-position with a methyl or ethyl group or a 3,5-dimethylpiperidino group, R _{3 is} 2-hydroxyethyl or 2-hydroxy-2-methyl-n-propyl and R _{4 is} 2-hydroxy-2-methyl-n-propyl, with or without a primary or secondary chemotherapeutic agent. into an effective drug against tumor resistance. 2,6-diethylmorpholino, cis-2,6-diethylmorpholino or 3,5-diethylmorpholino, R _{3 is} 2-hydroxyethyl, 2-hydroxy-n-propyl, 2-hydroxyisopropyl, 3-hydroxypropyl, 4-hydroxy-n-butyl, 2-hydroxy-2-methyl-n -propyl, 5-hydroxy-n-pentyl or 6-hydroxy-n-hexyl, methyl, ethyl, n-propyl or isopropyl, R ₄ 2-hydroxyethyl 2-hydroxy-n-propyl, 3-hydroxypropyl, 4-hydroxy-n-butyl or 2-hydroxy-2-methyl-n-propyl. 2. The method of claim 1, wherein the chemotherapeutic agent of natural origin is vinblastine, vincristine, vindesine, etoposide, teniposide, dactinomycin, daunorubicin, doxorubicin, bleomycin, mitramycin or mitomycin. 3. A process according to claim 1, wherein the compound of formula (I) is used R ₂ and R ₇ are independently selected from 2-methyl-piperidino, 3-methyl-piperidino, 4-methylpiperidino, 2-ethylpiperidino, 3-ethylpiperidino, 4-ethylpiperidino, 3,5 dimethylpiperidino, 3,5-diethylpiperidino, 2-methylmorpholino, 2-ethylmorpholino, 3-methylmorpholino, 3-ethylmorpholino, 2,6-dimethylmorpholino, cis-2,6-dimethylmorpholino, trans-2,6-dimethylmorpholino, 4. A process according to claim 1 wherein n is 0, R ₂ and R ₇ are independently selected from 3,5-dimethyl-piperidino, 2-methyl-morpholino, 2,6-morpholino, dimethylmorpholino cisz2.6- or trans-2,6-dimetilmorfolinocsoport, R _{3 is} methyl ethyl, 2-hydroxyethyl, 2-hydroxy-n-propyl, 2,3-dihydroxy-n-propyl or 2-methyl-2-hydroxy-n-propyl, and R ₄ 2-hydroxyethyl, 2-hydroxy-n-propyl or 2-methyl-2-n-propyl.