CH636616A5 - Process for preparing 7-substituted 7H-pyrrolo[3,2-f]quinazoline-1,3-diamines - Google Patents

Abstract

Novel 7-substituted 7H-pyrrolo[3,2-f]quinazoline-1,3-diamines of the formula <IMAGE> in which Y<2> denotes 3-aminobenzyl, 2-aminophenyl or 4-aminophenyl, as well as the non-toxic acid addition salts of these compounds, are prepared. The compounds of the formula I are obtained by reducing corresponding compounds in which Y<2> denotes 3-nitrobenzyl, 2-nitrophenyl or 4-nitrophenyl. The novel compounds may be used for preventing the growth of bacteria and as agents possessing an anti-malaria effect; they are active against lymphocytic leukemia P-388.

Description

The present invention relates to a process for the preparation of new 7- (substituted pyrrolo [3,2-f] -quinazoline-1,3-diamines. The compounds mentioned are biologically active.

Various derivatives of 2,4-diaminoquinazoline and 2,4,6-triaminoquinazoline are described in the literature and are known to have antifolic activity in bacterial systems. These compounds are also known to be antibacterial or antiprotozoal active. For example, 2,4-Diaminoquinazolines which have an alkyl group in the 5-position and / or 6-position or have a trimethylene bridge between the 5- and 6-position have an antibacterial activity [see Hitchings et al., U.S. Patent 2,945,859 or De Graw et al., J. Med. Chem., 17, 762 (1974), 2,4-diamino-6 - [(arylmethyl) amino] quinazoline; 2,4-diamino-6- ( [(substitutedearyl) methyl] arnino} -quinazolines; and 2,4-diamino-6 - ([(heterocyclic) methyl] amino} -quinazolines as well as the corresponding derivatives which have a 5-alkyl substituent or an N6 Alkyl substituents are active against malaria, [see Davoll et al., J. Med. Chem., 15.812 (1972); Eislager et al., J. Med. Chem., 15.1138 (1972); see likewise the newly edited article by E. Elslager "New Perspectives on the Chemotherapy of Malaria, Filariasis, and Leprosy", Progress in Drug Research, 18.99-172 (1974), in particular pages 111-116 and 152-154],

The [3,2-f] quinazoline-1,3-diamines which can be prepared according to the invention differ from the known 2,4,6-triaminoquinazolines in that they are linked by an ethylene group in the 5-position and in the N6 position, so that they form new tricyclic heterocyclic compounds.

The compounds which can be prepared according to the invention have the following formula:

H,

2nd

wherein

Y2 is 3-aminobenzyl, 2-aminophenyl or 4-aminophenyl.

The process according to the invention for the preparation of the new compounds of the formula I is characterized in that a compound of the formula

NH,

N-Y

wherein

Y3 means 3-nitrobenzyl, 2-nitrophenyl or 4-nitrophenyl, reduced and, if appropriate, compounds obtained converted into the corresponding salts.

The compounds of formula I and the non-toxic acid addition salts thereof, wherein

Y2 is defined above to prevent bacterial growth in vitro, as demonstrated in a standard tube dilution test using Saatagar or Wellcotest Sensitivity Test Agar amplified with 5% hemolosed horse blood as a wax medium. The compounds mentioned are active in vitro against one or more strains of the following bacteria: S. aureus smith, S. aureus 53-180, N-catarrhalis 8193, E. coli 9637, S. paratyphi 11737, K. pneumoniae 10031, or P vulgaris 6896. When the tube dilution test described above was carried out, the compounds gave MIC values in the range from <0.0009 5 / ml to 250 8 / ml compared to the test organisms.

In addition, the compounds of the formula I show an antifolic acid activity in vitro, as was shown by the inhibition of the growth of Streptoccus faecalis ATCC 8043 in a folic acid medium.

The compounds of formula I mentioned increase the antibacterial effect of "sulfa drugs". If you e.g. The following compounds were administered orally to mice to achieve a synergistic effect together with sulfamethoxazole against bacterial infections.

In addition, the above-mentioned are according to the invention

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The compounds of formula I are also active against malaria in vivo, as was shown by a standard blood schizonticidal test in mice infected with Plasmodium berghei KBG 173.

The activity was also demonstrated in vivo against P. cynomolgi infections in Rhesus monkeys, examining the following compounds: 7 (phenylmethyl) -7H-pyrrolo [3,2-f] quinazoline-1,3-diamine; 7 - [(4-methoxyphenyl) methyl] -7H-pyrrolo [3,3-fjquinazoline-1,3-diamine; and 7- [2,5-dimethylphenyl) methyl] -7H-pyrrolo- [3,2-f] chinazolin-1,3-diamine. The CDo doses of the compounds mentioned are 0.1 mg / kg / day, 0.316 mg / kg / day or 1.0 mg / kg / day when administered orally for 7 days.

Activity against P. berghei strains resistant to chloroquin, sulfones, cycloguanil and pyrimethamine in mice is demonstrated by using 7- (phenylmethyl) -7H-pyrrolo [3,2-f] quinazoline-1,3- diamine and 7 - [(2,5-dimethylphenyl) methyl] -7H-pyrrolo- [3,2- f] quinazoline-1,3-diamine were examined in mice against the resilient strains.

Certain compounds also have in vivo activity in mice (IP) against lymphocyclic leukemia P-388 when tested by the method described in Cancer Chemotherapy Reports, Vol. 3, No. 2, page 9 (Protocol 1,200) September, 1972 is described.

Any reducing agents which have the ability to reduce an aromatic nitro group to an aromatic amino group and which have no effect on other functional groups of the molecule can be used as the reducing agent in the process according to the invention. The preferred reducing agents are hydrogen in the presence of a noble metal catalyst, e.g. Palladium-Coal. An atmosphere pressure is preferred.

The compound of formula I obtained according to the invention can be isolated and purified, either in the form of the free base or in the form of the acid addition salt. Methods of converting one form to another are known to those skilled in the art.

For pharmacological uses, the compounds of formula I can be administered in the form of acid addition salts of a non-toxic organic or inorganic acid. These salts can be prepared by known methods. Preferred salts are those which are prepared with the acids mentioned below: hydrochloric acid, hydrobromic acid, maleic acid, benzoic acid, pamoic acid, methalsulfonic acid or acetic acid.

For pharmacological use, the compounds of formula I can be administered either alone or together with pharmaceutically acceptable carrier materials, the proportions and also the type of carrier being determined by the solubility and the chemical properties of the selected compound, also by the chosen route of administration and pharmaceutical practice. So e.g. compounds of formula I can be administered orally in the form of solid dosage forms, e.g. Capsules, tablets or powders, or in liquid forms such as e.g. in solutions or as suspensions. The compounds mentioned can also be administered in the form of injections by the parenteral route or in the form of sterile solutions or suspensions.

Solid oral forms of administration can include conventional additives such as e.g. contain the following: lactose, succrose, magnesium stearate, resins and similar materials. Liquid oral forms usually contain several

Flavors, colors, preservatives, stabilizers that simplify solubility or suspending agents. Parenteral preparations are usually sterile aqueous or non-aqueous solutions or suspensions containing various preservatives, stabilizers, buffer substances, solubilizers or suspending agents. If desired, additives can be added, e.g. Saline or glucose so that the solutions to be administered are isotonic.

Preferred examples of the method according to the invention are shown in the examples below. All temperatures are given in ° C.

example 1

7 - [(3-aminophenyl) methyl] -7H-pyrrolo [3,2-f] quinazoline-1,3-diamine

A mixture of 7.3 g of 7 - [(3-nitrophenyl) methyl] -7H-pyrrolo [3,2-f] quinazoline-1,3-diamine in 500 ml of acetic acid and 0.7 g of 10% palladium- Carbon catalyst is hydrogenated at atmospheric pressure at a temperature of 25 ° C. After about 2.5 hours, the hydrogen absorption ceases and the reaction mixture is filtered. The filtrate is freed from the solvent and the residue is stirred in an excess of an aqueous potassium carbonate solution, separated, it is washed with water and dried. This material is then recrystallized twice from methanol and dried to give 3.55 g of the triamine.

A solution of 3.4 g of the triamine in 50 ml of N hydrochloric acid is diluted with 100 ml of acetone and then cooled. The salt is separated off, it is washed with acetone and then dried, giving the compound mentioned in the title in the form of the dihydrochloride, monohydrate, salt, with a melting point of 328 ° C. with decomposition.

Analysis for: CnHi6N6.2HCl.H2O

Calc .: C 51.65; H 5.10; Cl 17.94; N, 21.26. Found: C, 51.40; H 4.77; Cl 18.22; N, 21.17

Example 2

7- (4-aminophenyl) -) h-pyrrolo [3,2-f | -quinazoline-1,3-diamine

A mixture of 3.20 g of 7- (nitrophenyl) -7H-pyrrolo- [2.3 f] quinazoline-1,3-diamine, 200 ml of glacial acetic acid and 0.5 g of a 10% palladium-carbon catalyst is used hydrogenated at atmospheric pressure and at about 25 ° C. After about UA hours the hydrogen uptake ceases, the mixture is filtered and the filtrate is freed from the solvent. The rest is recrystallized twice from water and then stirred with an aqueous potassium carbonate solution. The precipitate obtained in this way is washed with water and dried and then recrystallized from acetone, giving 1.48 g of the title compound which has a decomposition point of 180 to 185 ° C. and softens at 140 ° C. The compound is in the form of a hydrate and contains Vi molecule of water per molecule of triamine.

Analysis for: Ci6Hi4NeO.0,25H2O

Calc .: C 65.18; H 4.96; N 28.51 Found: C 65.34; H 4.92; N 28.37

Antibacterial activities

The ability of the compounds of formula I to prevent bacterial growth in vitro is demonstrated in the following test procedure:

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Prepare a stock solution or suspension of the test compound at a concentration of 2500 µj.m / ml by using a suitable solvent or medium such as e.g. aqueous sodium hydroxide, aqueous lactic acid, methyl cellic acid, dimethyl sulfoxide, dimethylacetamide, ethylene glycol, dimethylformamide, formamide, propylene glycol, acetone or methanol. Two-fold dilutions are made by adding appropriate amounts of sterile water to the solution or suspension of the test substance. One milliliter of each dilution is placed in seed agar or Wellcotest Sensitivity Test Agar, which is amplified with 5% hemolyzed horse blood (9 ml vol.) And is in sterile Petri vessels, giving plates containing different concentrations of the Test connection included. The hardened surfaces of each plate are incubated with the test organism and the plates are incubated at 35 ° C for 18 hours. The antibacterial activity in vitro of the investigated compounds is expressed as “minimum inhibitory concentration” (MIC), which defines the smallest amount of material (| im / ml) that completely inhibits the test organisms.

The antibacterial activities in vitro of the compounds obtained according to the invention are given in the following io tables, which contain the MIC values of the various compounds tested, which were examined by the method described above.

Table I

In vitro antibacterial activities of 7-substituted methyl-7H-pyrrolo [3,2-f-] quinazoline-1,3-diamine ii2n

n-ch2r

Compound R of examples

S. aureus Smith

S. aureus N. catarrhalis E. coli 53-180 8193 9637

MIC (y / ml.)

S. paratyphi IC. pneumoniae P. vulgaris "" "" 10031

11737

6896

Mediuma

1 3-amino- 0.488 0.244 0.016 0.244 0.976 0.122 1.95

phenyF

B

1 growth medium: A = seed agar with 5% hemolyzed horse blood.

B = Wellcotest Sensitivity Test Agar with 5% hemolyzed horse blood.

Table II

In vitro antibacterial activities of 7-substituted-7H-pyrrolo [3,2-fjchinazolin-1,3-diamimines

Compound R from example

S. aureus Smith

S. aureus 53-180

N. catarrhalis 8193

E. coli 9637

No

MIC (7 / ml.)

S. paratyphi K. pneumoniae P. vulgaris 11737 10031 6896

Mediuma

2 4-amino- 0.488 0.244 0.0152 0.244 0.488 0.031 0.976 B

phenyl a growth medium: A = seed agar with 5% hemolyzed horse blood.

B = Wellcotest Sensitivity Test Agar with 5% hemolyzed horse blood.

Antibacterial properties

The ability of the compounds obtainable according to the invention to have a synergistic effect against antibacterial infections in mice is demonstrated when these sulphomethoxazole are administered by the following test method:

The test agents are weighed, suspended in 0.5% aqueous carboxymethyl cellulose, homogenized (glass tissue crusher) and diluted accordingly, depending on the experiment to be carried out. Mice (male 18 ± lg.,

CD-1 strain) were weighed, distributed, randomly infused intraperitoneally with 0.5 ml of a standardized suspension (LDss ± 5%) of the bacterial organisms in 5% gastric mucin and randomly with individual doses of the test agents either during the Treated time of infection or 6 hours after infection. The treated groups consist of 10 mice per dose level. The dead mice were registered daily for 14 days and the PDso (the mice are treated at the time of infection) and the CDo (the mice

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are treated 6 hours after infection) are carried out according to the method of Reed and Muench [Amer. J. Hyg., 27,493 (1938)].

Antimalarial effects

The antimalarial effects of the compounds of the formula I obtained according to the invention are demonstrated and shown by the following test method:

Young ICR / HA Swiss mice and a standard inoculum from Plasmodium berghei KBK 173 are used. It is possible to cause a disease which is fatal to 100% of the untreated animals within 6-8 days, the mean survival time being 6 , Two days. The test animals weigh 18-22 g, but there may also be weight differences in any experimental or control group of 2-3 g. All animals were subjected to all tests and were approximately the same age. The animals that were tested were in plastic cages with a metallic top layer and they were given a standard laboratory diet and water ad libitum.

Intraperitoneal injections from 0.5 ml of a 1: 100 dilution of heparinized cardiac blood were administered to the depths with a minimum of 90% cells containing parasites (4 x 107 cells). These infected cells were obtained from donor mice that had been infected with Plasmodium berghei a week before. The effectiveness of the donor strain is maintained by weekly runs in various groups of mice inochulated with 0.5 ml of a 1: 500 dilution of heparinized heart blood.

The test compounds are administered as a solution or as a suspension in peanut oil. A single dose is administered subcutaneously after 72 hours where the mice have been infected with Plasmodium berghei. At this point 10-15% of the parasites had developed; the disease could be recognized well, but there was no way to develop sufficient weakness to change the guest's response to the toxic effects of the drug being tested. Since the treatment was carried out for 3 days so that the infection could “nest” well and dead animals appeared in controls within 6-8 days in the untreated animals, it is assumed that this system is a preferred connection with a maximum Represents requirement. To determine such factors as e.g. Changes in infection with Plasmodium berghei or in the sensitivity of the guest or to find technical errors, a group of infected animals were treated with pyrime-io thamine at dose level, which produced certain increases in survival time, with each attempt a positive control includes.

In each experiment, the test compounds were administered in graduated dosage forms. With highly active compounds, an increased dose level subsequently increases the survival time of the treated mice. However, if the active drug is toxic to the guest, this toxicity can be a limiting factor; Continuous increases in the dose level also increase the toxic effects and can shorten the survival time. Deaths before the 6th day when uncontrolled animals start to die are considered non-parasitic and are the basis for the toxicity evaluations. Treated animals are observed for 60 days. Surviving animals after this period are considered

that they are healed. When calculating survival time, toxic deaths and 60-day survivors are not included.

The compound obtainable according to the invention is considered active if at least one of the test animals is cured or if a significant increase in the survival time of treated animals is found compared to the survival time of untreated animals, but with the proviso that the medicaments administered do not Death occurs when administered in amounts of the active dose (toxicity).

The results of the antimalaria tests of the compounds of the formula I obtained according to the invention are summarized in the tables below.

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Table III

Activity of 7- (substituted) methyl-7H-pyrrolo [3,2-J] quinazoline-1,3-diamines against Plasmodium berghei KGB 173 malaria in mice (all doses are in mg. Per kg, see above)

V

n-ch „b

Connection of R

example

Highest non-lethal lowest dose, the min

Dosea heals all mice (healing / treatment)

Can

1 3-aminophenyl 80 20 5 (3/5) 7.8 1.25

Attempted activity

Individual doses of 7- (phenylmethyl) -7H-pyrrolo- [3,2-f] quinazoline-1,3-diamine (active part) are administered intraperitoneally in 0.5% carboxymethyl cellulose at a constant dose. Volume of 10 ml / kg administered to groups of 10 mice per dose level. (Charles River COBS CD strain albino mice, male, weight 21.0-25.29).

All mice were observed daily for the duration of the experiment, 14 days. The LD 50 obtained was 54.5 mg / kg (95% confidence limits of 48.6- "s 93.4), and all deaths occurred within 4-5 days after drug administration.

The compound obtainable according to the invention spontaneously reduce motor activity, bradypnea, ptosis and

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Expansion with the sides drawn in all mice within one hour after the injection. In addition, rough coatings and a reduced fecal excretion were observed on the 2nd day after the administration of the drug. These effects lasted up to day 6 in animals receiving the low doses and 14 days in those receiving the highest doses. All mice that were still alive on day 14 were sacrificed and examined. Macroscopic examinations have shown that the tissues are in their normal condition.

B

Claims (2)

636 616 2nd PATENT CLAIMS 1. Process for the preparation of a new compound of the formula NH. wherein Y2 is 3-aminobenzyl, 2-aminophenyl or 4-aminophenyl and the non-toxic acid addition salts of these compounds, characterized in that a compound of the formula nh, wherein Y3 means 3-nitrobenzyl, 2-nitrophenyl or 4-nitrophenyl, reduced and, if appropriate, compounds obtained converted into the corresponding salts. 2. The method according to claim 1, characterized in that hydrogen is used as the reducing agent in the presence of a noble metal catalyst.