CS207700B2 - Method of makingthe new 2-aryl-1h-perimidine compounds - Google Patents

Description

The present invention relates to a process for the preparation of novel 2-aryl-1H-perimidine compounds and pharmaceutically acceptable salts thereof.

Previous 2-eubstituted-1H-perimidine compounds and 2-substituted-2,3-dihydro-1H-perimidine compounds have been described which differ from the compounds of the invention.

Article in Why. Am. Assn. Cancer Res. J, 319 (1962) reports on the carcinostatic activity of 2- (3,4-dichlorophenyl) -1H-benzo [d] quinazoline, the same compounds as 2- (3,4-dichlorophenyl) -1H-perimidine. The article explicitly states that the compound is ineffective when administered orally.

Article in J. Het. Chem. J, 108 (1964) is the following study inspired by the aforementioned article. A series of 2-aryl-2,3-dihydro-1H-perimidines was synthesized in this study. In this series were:

2- (p-chlorophenyl) -2,3-dihydro-1H-perimidine

2- (3,4-dichlorophenyl) -2,3-dihydro-1H-perimidine

2- (p-bromophenyl) -2,3-dihydro-1H-perimidine.

The work says nothing about any usefulness of these compounds.

However, the usefulness of the compounds of the above series is described in J. Med. Chem. 9 (4),

599 (1966). The three compounds tested had little antineoplastic activity.

Article in <J. Org. Chem. 36, No. 11,1477 (1971) discloses, inter alia, 2-substituted-1H-perimidines and their p-toluenesulfonate salts including 2-phenyl-1H-perimidine, 2- (p-chlorophenyl-1H-perimidine), and 2- (p-methylphenyl) -1H-perimidine and their p-toluene sulfonate salts This citation says nothing about their usefulness Similarly, the article in Reaction Spoeobnost, Org. Soedin, 6 (1) 47-54 (1969) (Eng.) describes 2-phenyl-1H-perimidine, 2- (p-tolyl) -1H-perimidine, 2- (p-bromophenyl) -1H-perimidine and 2- (m-bromophenyl) -1H-perimidine, but do not state their applicability .

The following references in principle relate to the production of 1,2-substituted perimidines.

Soviet Patent No. 504,770 reports on perimidines that have biological activity and could find use in medicine, but there are no notes or references that authenticate this comment, which moreover applies only to 1,2-dieubettulated compounds.

Khim. Parm. Zhur., 11 (5) 87-93 (1977) describes the efficacy of 1,2-disubstituted perlmidines and 2- (3,4-dimethoxyphenyl) perimidine on the central nervous system.

The compounds of the invention distinguish them from the compounds in these publications in that they are unsubstituted at the 1-position and the compounds produced by the process of the invention have additional sub-substituents on the phenyl ring. In addition, the invention is directed to suppressing an immunological response in a mammal, which is different from central nervous system activity or cancerostatic activity.

The present invention provides a process for the preparation of the novel 2-aryl-1H-perimidine compounds of formula I

HN-R

where

R is a radical of formula

NH where

R ¹ represents -CF, -OCFj, -SCFj -OCgFij or town or in the position para and pharmaceutically acceptable salts thereof, characterized by condensing 1,8-lan diamlnonafta IV

NH ₂ NH ₂ (IV) with an acyl compound of formula V wherein

R ¹ is as defined above.

(IN)

In formula I, the carbon atom of the group of formula

NH is always in the middle between 2 nitrogen atoms, so formula I determines only one type of structure (III) where

My ^R1 as defined above.

Similar substances are described in J. Het. Chem. J., 108 (1964), J. Org. Chem. 36 (12),

477 (1971) and references cited in both articles.

Most of the compounds of formula V which are used as starting materials are known compounds and all are prepared by conventional methods, m- and p-trifluoromethoxybenzoyl fluorides are prepared by chlorination of alkyl m- or p-methoxybenzoate and reacting the formed p-trichloromethoxybenzoyl chloride with SbF. -j or SbCl 2. m- and p-trifluoromethylthiobenzoyl chlorides are prepared by reaction with an alkyl m- or p-iodobenzoate with Hg (SCF ₃ ) ₂ , hydrolyzing the resulting alkyl m- or p-trifluoromethylthiobenzoate and converting the free acid to an acid chloride by treatment with, for example, SOCl ₂ . The corresponding aldehydes used to prepare the compounds of formula III are prepared from the acyl halides by conventional methods.

The compounds of formula I are useful for suppressing immunological reactions in mammals. Such suppression involves suppressing the immune response induced whenever the mammalian body generates antibodies and reactive cells in response to the presence of a foreign protein (protein). The practical application of immunosuppressive protein is diverse. An important application of immunosuppressive activity is organ transplantation, but the immunosuppressive effect can advantageously be used in the treatment of various diseases collectively referred to as autoimmune diseases. Autoimmune diseases include autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, lupus erythromatoeus, lupoid hepatitis, lupus nephritis, glomerulonephritis, nephrotic syndrome, Goadpastr's syndrome, Wegener's granulomatosis Psoriasis, scleroderma, scleroderma, scleroderma .

The compounds of formula I can be administered both orally and parenterally. The exact amount of active ingredient to be used varies with the individual compounds. However, the compounds have a high therapeutic index, so that effective, non-toxic doses have a wide range in each case. Depending on the test system, this range for the more efficient members of the test series ranges from less than 1.6 mg / kg / day to 25 mg / kg / day. The other compounds of the series require larger doses of up to 100 mg / kg / day or more in small mammals. The given dose relationship for small and large animals described for other substances, for example, the human dose of the immunosuppressive agent azathrioprine is usually 1-2 mg / kg, while for the mouse it is approximately 50 mg (see also Cancer Chemotherapy Reports 50: 219). 1966/1, the predicted effective human doses should be correspondingly lower than in small mammals, from 0.5 to 10 mg / kg / day.

The compounds of formula 1 are best administered as pharmaceutical preparations. Pharmaceutical compositions are well known in pharmaceutical practice. When preparing preparations with current

207700 4 active compounds of the formulas, the selected compound is mixed with a carrier such as lactose, dextrose, sucrose, sorbitol, aannitol, starches, acacia rubber, calcium phosphate, alginates, tragacanth, gelatin, methylcellulose, starch, magnesium atearate or mineral oil. The compositions may be formulated as tablets, suspensions, or capsules. For parenteral use, the compositions are formulated as injectable solutions.

A preferred formulation is one which, in a single dosage form adapted for psroral administration to obtain an immunosuppressive effect, an immunosuppressive, nontoxic amount ranging from 10 to 1,000 mg of active ingredient · pharmaceutical carrier.

The invention will be illustrated by the following example.

Example 1

Preparation of 2,3-dlhydro-2- (p-trifluoromethylphenyl) -1H-psrimidine

A solution containing 15.8 g (0.10 mol) of 1,8-diaminonaphthalene, 17.4 g (0.10 mol) of p-trifluoromethylbenzaldehyde and 1 liter of xylene was heated at reflux for 24 hours. The solvent was evaporated in vacuo to give 37.5 g of crude product which was purified on a toluene-eluted silica gel column to give the title compound (22.93 g, 73%), mp 1-9-122 ° C. . Mass Spectrum m / e 314.

Analysis: Calculated: C 68.78; H 4.17; N 8.91 56;

Found:% C, 68.69;% H, 4.40%;

Example 1a

A process for the preparation of 2,3-dihydro-2- (p-trifluoromethoxyphenyl) -1H-periaidine

8.0 St LJ. 0.0506 moles of 1,8-diaminonaphthalene and 9.62 g, i.e. 0.0506 moles of p-trifluoromethoxybenzaldehyde, were reacted as in Example 1 to give 5.76 g of the title compound after recrystallization in 34.4% yield. mp 103-105 ° C, mass spectrum m / e 330.

Example 1b

A process for the preparation of 2,3-dihydro-2- (p-trifluoromethylthiophenyl) -1H-perimidine

4.68 g (0.0296 mole) of 1,8-diaminonaphthalene and 4.98 g (0.0296 mole) of p-trifluoromethylthiobenzaldehyde are reacted as in Example 1 to give 5, (53.2% yield). 45 g of the title compound, m.p. 133-134 ° C, mass spectrum m / e 346. Anal. Calcd. For C, Η ^ ^NgSF ₃ : C, 62.42; C, 3.78; 56; H, 8.09; % H, 4.01;% N, 8.06.

Hemagglutinin test in mice by oral administration

Groups of 5 Swiss male mice weighing 20 g each are separated and given intravenously 5 x 10 sheep red blood cells. Blood cells for this administration are prepared from sheep's blood in Alsever's solution, then washed three times with 0.85% sodium chloride and resuspended in the same solution. Orally, 0.1 ml of the test substance in solution in polyethylene glycol 400 is administered orally at a dose of 0.1 ml, starting from 3 days before the injection of red blood cells. Multiple doses are used for each compound, each additional dose being twice the previous dose. The control group of mice receives only the injection of red blood cells and orally only the solvent instead of the active ingredient. Six days after antigen injection, mice are bled and sera from each group of 5 mice are mixed. Following inactivation of the compliment, the serum hemagglutinin content is determined by standard procedures using a dilution mixture in sodium chloride containing the serum of interest and a 0.5% suspension of sheep red blood cells. Each subsequent dilution has half the content of said substances compared to the previous dilution. The tests are carried out on plastic supports. The solutions were incubated for 3 hours at 37 ° C on which plates the hemagglutination degree was determined. If more than four-fold (75%) or greater antibody decreases occur in the serum of interest compared to the control serum, the result is considered statistically significant. The results are expressed as the minimum effective dose (MED), which is the lowest dose that causes the antibody content to be reduced to 75% or greater.

The results are shown in Table 1 below: (MED is the minimum effective dose, reducing antibody production by at least 75% when administered orally to PO).

Table I

Compound MED mg / kg x9 PO

2,3-dihydro-2- (p-trifluoromethylphenyl) -1H-perimidine 25

2,3-dihydro-2- (p-trifluoromethoxyphenyl) -1H-perimidine 6.25

2,3-dihydro-2- (p-trifluoromethylthiophenyl) -1H-perimidine 12.5

Determination in individual sera

During these experiments, the previous procedure was modified so that 10 mice were used per group instead of the original 5 mice. Mice were bled as above but the antibody content was determined in individual sera instead of in the serum mix. The mean hemagglutinin (log ₂ ) + SE value and p value were calculated for each group of 10 mice by Student's test. The results were compared with the control group. A dose of the active compound that statistically significantly (p &lt; 0.05) reduced the antibody titer was sought. The active compounds were administered orally or subcutaneously for 10 days starting from 3 days prior to administration of the red blood cell suspension in a solution consisting of sodium chloride containing 0.125% methylcellulose and 0.2% nonionic emulsifiers. The assay was performed 7 days after the injection of blood cells.

Graft Host Response (GVH)

In this assay, mouse spleen cells (C57BL) were administered parenterally to mice of hybrid strain F, (C57BL x C3H). In this case, the mice to which the spleen cells are administered do not reject these cells because the hybrid strain recognizes the related C57BL antigen as its own. However, the cells thus administered increase the response of recipient tissues to C3H-derived antigens. As a result, the spleen of the recipient becomes larger. If an immunological response is prevented, this splenic enlargement will not occur. For this reason, spleen weight can be used as a measure of GVH due to suppression of the immunological response.

Modifications to the original Simons procedure were used in the experiments (Ann. N.Y. Acad. Sci. 73: 834, 1958). In this way, a larger number of spleen cells can be obtained using a Waring blandors. In this method of obtaining spleen cells, the process proceeds completely automatically. Double homogenization is performed after 6 seconds using 25 C57BL spleens in 23 ml of sodium chloride solution. This procedure is sufficient to release the cells from the connective tissue. This tissue is removed by filtration through several layers of mule. The cell suspension prepared in this way is standardized in a Levy-Hausser chamber to contain 6 x 10 6 cells with a core per ml. Thereafter, groups of 10 C57BL x C3H mice weighing 16 to 18 g were administered this suspension intraperitoneally with a total content of 1 ml. The active substances are administered orally or subcutaneously in a volume of 0.2 ml, starting 3 days prior to the cell injection and then daily for 13 days. Control cells are given only cells and solvents. The spleens are removed and weighed 10 days after cell injection. Results are expressed as spleen weight / g body weight.

These injections cause a small enlargement of the spleen, and the weight of the spleen can be used to determine 100% suppression of the GVH response. The calculation method is given in the following example for mice treated with a comparator compound suppressing an immunological reaction.

Treatment with control compound mg spleen / g body weight + SE ^X ) % inhibition ") 1- (6-methoxy-2-benzothiazolyl) -3-phenylurea (J. lled. Chem. 12, 016-1018 (1969)), 12.5 mg / kg x 13 (oral) 6.86 i 0.80 ^xxx ) 74 none (GVH control) 11, 55 ± 1.01 0 none (syn. check) 5.20 and 0.37 100 ALIGN! none (normal check) 4.16 and 0.17 -

^x ) Mean of groups of 5 mice (GVH control - treated mice) ^xx ) - x 100 =% inhibition (GVH control - syn control) ^xxx ) p <0.01 compared to GVH control

Since in practice, it was found that the syngeneic and normal controls differ very little in different experiments, the total value (4.8) was used for the calculations, which was derived from repeated calculations using four different syngeneic control groups ( 5.20 ± 0.37, 4.99 ± 0.39, 4.42 ± 0.13, 4.66 ♦ 0.12) and were considered to comprise 20 mice.

Arthritis test using adjuvant in rats

2- (p-Trifluoromethylphenyl) -1H-perimidine hydrochloride was examined for its ability to swell the hindpaw and bone damage due to swelling induced by adjuvant administration in rats. In order to quantify the inhibition of swelling, two stages of inflammation have been defined:

(1) primary and secondary hindpaw inflammation injected; and (2) secondary hindpaw inflammation injected. This inflammation usually develops 9 days after induction of inflammation in the injected paw. A decrease in the second type of inflammation indicates the suppression of the immunological response as described by Chang, Arth. Rheum. 20,

135-1111 (1971). Phenoprofen at 30 mg / kg was used as a standard anti-inflammatory compound for comparative evaluation. (Nickander et al., Med. Proc. Annual FASEB Mtgs., April 1971, ABS # 205.)

Arthritis of the above type was induced in male Lewis-Wistar rats weighing 200-210 g by administering a single subplantar injection to the right hind paw of the male, containing 0.1 ml of a 0.5% suspension of heat-killed, lyophilized Mycobacterium tuberculosis (Calbiochem. Perrigen-C) in mineral oil. The procedure is a modification of the method of Winter et al., Arth. Rheum. 9: 394-397 (1966). One group of 5 rats (TB control) received only this injection. Further, a group of 5 rats were left without any treatment (normal controls). Each of the compounds was suspended in a 1% suspension in carboxymethylcellulose and administered by gavage to groups of 5 rats per day at a dose of 30 mg / kg from day 1, 7 days after adjuvant injection. Paw volume was measured using a digital voltmeter and a Statham pressure transducer to transmit pressure values. Both hindpaw volumes were measured on days 2, 4, 7, 9, 11, 14, 16 and 18. X-ray images were obtained on day 18 after sacrifice of the animals. Non-injected paw volumes were measured from day 9 to day 18 and are listed in the following table.

Table

Treatment Paw volume that was not injected from day 9 to day 18 day 9 day! 1 day 14 day 16 day 18 (normal

checks) 1.99 and 0.063 2.13 i 0.08 2.08 í 0.053 2.15 ± 0.056 2.01 í 0.95 TB checks 1.94 0,0 0.028 2,02 í 0,037 2.69 í 0.106 3.26 and 0.202 3.65 í 0.227 phenoprofen 1.97 and 0.023 2.11 i 0.07 2.38 í 0,099 2.7 ± 0.107 2.67 i 0,128 " (23% inhibition) *)

volume of treated animals - volume in normal controls ^x )% inhibition = 1 - x 100 volume in TB controls - volume in normal controls

Examination of the injected or non-injected paw X-ray showed that there was no bone damage in the animals treated with 2- (p-trifluoromethylphenyl) -1H-perimidine hydrochloride, but this was very evident in TB controls.

Claims (2)

SUBJECT OF THE INVENTION A process for the preparation of the novel 2-aryl-1H-perimidine compounds of the general formula I NH R is a radical of formula wherein R ¹ is -CF 3, -OCF 3, or -SCFj -OCgF ^ in meta or para position and their pharmaceutically acceptable salts, characterized by condensing 1,8-diaminonafta only formula IV NH ₂ NH ₂ (IV) with an acyl a compound of formula V (V) wherein R ¹ is as defined above. 2. A process according to claim 1 for the preparation of 2,3-dihydro-2- (p-trifluoromethylphenyl) -1H-perimidine, characterized in that 1,8-diaminonaphthalene is reacted with p-trifluoromethylbenzaldehyde.