RU2146927C1 - Preparation for treatment of patient with cancer skin and precancer skin diseases - Google Patents

Abstract

FIELD: medicine, oncology, pharmacy. SUBSTANCE: invention proposes glyciphon ointment containing lanolin, Vaseline, emulsion waxes and methylphosphonic acid diglycidyl ester of the chemical formula:

Description

 The invention relates to medicine, to drugs for the treatment of cancer and precancerous skin diseases, namely, a new drug and its dosage form.

 It is known that for the treatment of these diseases, 0.5% colchamin ointment, 25% ointment with podophylline, Gordeev’s liquid N 2, Karchauli liquid were used, but due to insufficient selectivity of action or high toxicity, they were not widely used.

 Currently, 30% and 50% prospidine ointment is used for the treatment of basal cell carcinomas, skin cancer and 0.5% colchamin ointment for the treatment of skin cancer of I and II stages. Colchamin ointment has a complex composition: in addition to colchamine, it contains antiseptic agents thymol and ethyl alcohol; as well as an antibiotic - synthomycin. It has a skin-resorptive effect, which is manifested by leukopenia and other side effects. When using a prospidin ointment, hyperemia, swelling, fibrinous deposits of surrounding healthy tissue are possible. (Mashkovsky M. D. Medicines, M., "Medicine", 1993). Closest to the claimed invention is a cytostatic drug containing as an active substance a compound with glycidyl groups and a pharmaceutical diluent prototype (DE N 3634745).

 However, the range of topical antitumor agents for skin cancer and precancerosis is limited.

 In order to expand the range of ointments with local anti-blastoma activity, the task was set - to create a new drug for the treatment of cancer and precancerous skin diseases.

Химическое соединение - диглицидиловый эфир метилфосфоновой кислоты известно и описано в литературе (Ризположенский Н.И., Бойко Л.В., Зверева М. А. "Синтез глицидиловых эфиров кислот фосфора". - Доклад АН СССР, 1964, - 155, 5, с. 1137-1139).The problem is solved in that the drug includes an active substance and a pharmaceutical base. As the active substance used diglycidyl ether methylphosphonic acid (Glyciphone) of the following chemical structure:

The chemical compound - diglycidyl ether of methylphosphonic acid is known and described in the literature (Rizpolezhensky N.I., Boyko L.V., Zvereva M. A. "Synthesis of glycidyl ethers of phosphorus acids." - Report of the USSR Academy of Sciences, 1964, - 155, 5, p. 1137-1139).

 The use of this compound in medicine is not described in the literature.

Its dosage form is an ointment based on lanolin, petroleum jelly and emulsion wax in the following ratios of components,%:
Methylphosphonic acid diglycidyl ether (Glyciphone) - 5 - 50
Anhydrous lanolin - 50 - 94
Vaseline - No more than 45
Emulsion Waxes - Not more than 10
The introduction of glyciphone below the lower limit does not provide a therapeutic effect, above the upper limit, the stability of the dosage form is violated.

 The introduction of anhydrous lanolin below the lower limit does not provide the function of the hydrophilizing and stabilizing component of the ointment base.

 The introduction of petroleum jelly improves the consumer properties of the ointment - gives it plasticity. The introduction of vaseline above the upper limit does not allow to obtain an aggregatively stable system.

 The introduction of emulsion waxes in the ointment promotes the hydrophilization of petroleum jelly and eliminates the appearance of phase heterogeneity of the surface and structure of the ointment. The introduction of emulsion waxes above the upper limit leads to their crystallization from the ointment.

 The name "Glyciphone" was assigned to the drug by the decision of the Nomenclature Commission of the Pharmacological Committee of the USSR Ministry of Health on January 24, 71 (Protocol No. 1).

 Compared with known drugs, the proposed ointment has significant advantages: it exhibits a high selectivity of action on blastomatous cells, combined with antimicrobial activity and harmlessness to surrounding normal tissues, as well as the absence of side skin-resorptive action. Its advantage is that the ointment can be used with a good cosmetic effect in the localization of tumors and precanceroses on the skin in the eyelids, back of the nose, auricle and external auditory canal, as well as its effectiveness in relapse of tumors after surgical and radiation treatment.

 The proposed tool was subjected to experimental and clinical studies. The ointment has passed successful clinical trials in leading oncology clinics in Moscow, St. Petersburg, Kazan and other cities.

 The high contact anti-blastoma activity in transplantable ascites tumors in mice and rats, its harmlessness to the hematopoietic system of animal tumor carriers, as well as the effectiveness of glyciphone ointment in cancer and precancerous skin diseases are confirmed by the following examples.

 Example 1. The effect of Glycifon on the growth of transplantable ascites tumors.

 Methodology

 The experiments were performed on 220 mice and 74 rats with ascitic transplant tumors. The ascites tumor strains that were widely used in experimental tumor chemotherapy were used: sarcoma 37, Ehrlich tumor, N K / Ly lymphoma, sarcoma 180, rat ovarian tumor. (Pogosyants EE, Kiseleva NS "Tumor strains supported at the Institute of Experimental and Clinical Oncology of the Academy of Medical Sciences of the USSR" - Questions of Oncology, 1963, 9, 8 - pp. 103-116).

 Tumor transplantation was carried out according to the generally accepted technique (Pershin G.N. Methods of experimental chemotherapy. - Medicine, M, 1971. - 539 p.).

 When the tumors were intraperitoneally inoculated with ascitic fluid, it was diluted with 4% sodium citrate solution in physiological saline so that 0.2 ml of diluted ascitic fluid contained 3 million tumor cells.

где K - объем асцитической жидкости в контроле,
O - объем асцитической жидкости в опытной группе.Treatment began one day after transplantation. Glyciphone in freshly prepared aqueous solutions was administered intraperitoneally daily to rats at 50 mg / kg, and to mice at 60 mg / kg for 10 days. During the experiments, the mass of mice and rats was recorded. After completing the course of treatment, the animals were decapitated under mild ether anesthesia, and the volume of ascitic fluid was determined. Comparing these values in the experiment and control, the percentage of inhibition of tumor growth was calculated by the formula:

where K is the volume of ascitic fluid in the control,
O is the volume of ascitic fluid in the experimental group.

 All experimental results were processed by the statistical method of indirect differences (Monceviciute-Ehringen E.V. Simplified mathematical and statistical methods in medical research. Pathological physiology and experimental therapy, 1964, N 4, pp. 71-78).

results
Evaluation of anti-blastoma activity showed that glyciphone inhibits the growth of ascites tumors by 80-100%, and in a significant part of the animals at the end of the experiment ascitic fluid was not found in the abdominal cavity (see Table 1).

Conclusion
Glyciphone exhibits high contact anti-blastoma activity in transplantable ascites tumors of rats and mice without a pronounced general toxic effect.

Example 2. The safety of intraperitoneal administration of glyciphone for the hematopoietic system of rats with ascites tumor
In a series of experiments on rats with an ovarian tumor, peripheral blood indices of experimental and control rats were compared. A blood test of 10 experimental (after a 10-day course of intraperitoneal injections of glycifon at 50 mg / kg) and 10 control (administration of distilled water) animals was performed according to the standard method.

 The research results are given in table. 2.

It was found that the number of leukocytes, erythrocytes, hemoglobin content and color indicator of the treated animals did not significantly differ from those of control rats. Assessment of the qualitative composition of leukocytes revealed an increase in the number of monocytes in the blood of experimental animals (see table. 2)
Conclusion
Glyciphone with 10-day administration to rats with an ovarian tumor of 50 mg / kg daily does not show a hematotoxic effect.

Example 3. The results of a clinical study of the effectiveness of topical application of glycyphonic ointment in the treatment of patients with skin cancer and basal cell carcinoma
Patient Characterization
445 patients with various clinical types of basal cell carcinomas and 20 patients with stage I squamous cell carcinoma were treated at the Department of Skin and Sexually Transmitted Diseases of Kazan Medical University. Among the patients were 176 men and 289 women aged 30 to 83 years.

 According to the clinical types, basal cell carcinomas were distributed as follows: tumor type - in 230 patients, ulcerative - in 9, superficial type - in 206.

 In the vast majority of patients, lesions were localized in the face, less often on the trunk and limbs. In 440 patients, lesions were single, in 5 - multiple. The disease duration in 100 patients was up to 1 year, in 230 - up to 3 years, in 135 - up to 5 years or more.

 The diagnosis was established based on the results of clinical, histological or cytological studies.

 Before treatment, patients underwent a clinical examination. In addition, before and at the end of treatment, a general analysis of blood and urine was carried out, cholesterol, prothrombin, bilirubin, and blood glucose were determined, and a proteinogram was studied.

Treatment technique
In most cases, patients were treated on an outpatient basis. Before applying the ointment, the skin in the area and around it was wiped with a gauze or cotton ball moistened with alcohol. A 30% glyciphon ointment of the following composition was applied to the tumor:
Methylphosphonic acid diglycidyl ether (glyciphone) - 30.0
Anhydrous lanolin - 60.0
Vaseline - 10.0
The dose of ointment depended on the size of the tumor - from 0.8 to 1.5 g - with the capture of the surrounding skin up to 0.5 cm and covered with a napkin of 4 layers of gauze. A piece of wax paper was placed between the two layers of the napkin so that the upper layers of the napkin did not soak the ointment. The napkin was fixed with a sticky patch. Applications of glycyphon ointment were used 1 time per day. At each dressing, necrotic tissue from the surface of the lesion was removed with a blunt scalpel.

Treatment results
Usually, after the first three applications of glyciphone ointment, some loosening of the tumor was noted. Around her there was a slight hyperemia. After 6 or more applications, the tumor was destroyed, a tissue defect appeared in the focus. Suppuration in places of applications was not noted in any case. For complete destruction of the tumor, 9 to 12 were performed, less often - 15 applications.

 After the termination of the applications of the glyciphon ointment, aseptic dressings were applied to the resulting defect before scarring, which occurred within 15-20 days. A slightly retracted, depigmented, smooth scar remained at the site of the tumor. Subsequently, the scar simplified and became invisible.

 In the treatment of various clinical types of basal cell carcinoma, it was noted that small-node forms (tumor type), when localized in the forehead and nose, are more resistant to glyciphone ointment therapy. Therefore, with these forms of basal cell carcinoma, the number of applications was adjusted to 15.

 As a rule, with other locations of the tumor, pronounced perifocal changes in the surrounding tissues were not observed.

 A feature of the use of glyciphon ointment in the localization of tumors in the orbital region is that after 5-6 applications, edema of varying severity was sometimes observed, which usually did not prevent the continuation of treatment. This edema, apparently, is explained by the peculiarity of the anatomical structure of this area.

 In the treatment of patients with skin cancer and basal cell carcinomas, glycyphon ointment did not reveal any pronounced side effects. The general condition of the patients was not impaired. The parameters of peripheral blood, urinalysis, and bilirubin, prothrombin, blood glucose, and proteinogram did not change.

 Of the 465 patients, 462 achieved persistent clinical recovery. Duration of observation from 1 year to 7 years. The reason for the failure of the primary treatment in 3 patients was the insufficient number of applications due to the fault of the patients themselves. All 3 patients were subsequently re-treated with glyciphone ointment with good results.

Conclusion
Glyciphon ointment is an effective therapeutic agent in the treatment of patients with various clinical and morphological types of basal cell carcinomas and squamous cell carcinoma. The treatment method is simple, affordable and recommended for wide clinical use.

 These treatment results are confirmed in the Oncology Research Institute named after N.N. Petrova and Moscow Clinical Research Institute. M.F. Vladimirsky.

 Example 4. Evaluation of the effectiveness of glyciphone ointments containing emulsion waxes.

According to the scheme shown in example 3, was treated 10 patients with basal cell carcinomas with glyciphon ointment, of the following composition:
Methylphosphonic acid diglycidyl ether (Glyciphone) - 30.0
Anhydrous lanolin - 59.0
Vaseline - 10.0
Emulsion Waxes - 1.0
Clinical observations showed that there was no significant difference in the nature of the action of the ointment containing emulsion waxes. In all 10 cases, a therapeutic effect and healing was obtained.

 The advantage of an ointment containing emulsion waxes is its homogeneity and aggregative stability during storage, which is confirmed by visual observations for 2 years and the determination of colloidal stability by centrifuging samples (for 5 minutes at 6000 rpm) for different periods of storage. At the same time, the addition of emulsion waxes does not significantly affect the release of glyciphon-substance from the ointment, which is confirmed by the results of special studies below.

 The ability of drugs to penetrate the skin can be judged by the results of an in vitro release study (Alyushin M.T. Influence of excipients on the bioavailability of drugs. - In the book: Problems of containers, packaging and auxiliary materials in pharmacy. - VNIIMI, M ., 1978, p. 53-56).

 The effect of emulsion waxes on the release of glyciphone from ointments by the equilibrium dialysis method according to Kruvchinsky through the shell shell of a chicken egg was studied.

Methodology
0.5 g of ointment (accurate weighed) was weighed onto a circle of parchment paper and placed in the dialyzer on the membrane with the paper up, evenly distributing the ointment over its entire surface. The dialyzer was placed in a beaker with a capacity of 100 ml, into which 30 ml of distilled water at a temperature of 37 ^° C was previously poured. The device was placed in a thermostat (37 ^° C ± 1 ^° C). After 0.5, 1, 2, 4, 6, 8, and 24 hours, samples were taken by completely replacing the receptor medium. Quantitative determination of glyciphon in the samples was carried out spectrophotometrically on SF-46 using phosphorus, according to the formation of a colored vanadate-molybdate-phosphorus complex after mineralization with concentrated sulfuric acid in the presence of potassium permanganate according to the procedure given in VFS 42-2277-93 for glyciphone substance.

где X - содержание глицифона в пробе, %;
D_x - оптическая плотность анализируемого раствора;
D₀ - оптическая плотность стандартного раствора;
0,000008 - содержание фосфора в рабочем стандартном растворе, г;
a - навеска мази, взятая для анализа, г;
14,89 ± 0,2 - содержание фосфора в глицифоне;
30 - объем диализной среды, мл;

Расчет количества глицифона, высвободившегося из мази, проводили по формуле

где X - количество глицифона, высвобожденного из мази,%;
n - порядковый номер пробы;
C_i - количество глицифона, высвободившегося из i пробы,%.The calculation of the quantitative content of glyciphone in the samples was carried out according to the formula

where X is the glyciphon content in the sample,%;
D _x is the optical density of the analyzed solution;
D ₀ is the optical density of the standard solution;
0,000008 - phosphorus content in the working standard solution, g;
a - a sample of ointment taken for analysis, g;
14.89 ± 0.2 - phosphorus content in glyciphone;
30 - the volume of the dialysis medium, ml;

The calculation of the amount of glyciphone released from the ointment was carried out according to the formula

where X is the amount of glyciphone released from the ointment,%;
n is the serial number of the sample;
C _i is the amount of glyciphone released from i sample,%.

 Statistical analysis of the obtained data was carried out by the method of GF XI.

 The results of the release of glyciphone from ointments are presented in table. 3.

From the data table. 3 it follows that the introduction of emulsion waxes in the composition of the ointment in an amount of 1% does not affect the rate and degree of release of glyciphone from the ointment
Example 5. Treatment of patients with precancerous skin diseases: senile keratoses and Boven’s disease.

Patient Characterization
There were 36 patients with senile keratoses, of whom 14 were men and 22 were women. Patients were distributed according to age as follows: from 40 to 50 years - 1, from 51 to 60 - 9, from 61 to 70 - 15, older than 70 - 11 man.

 Disease duration up to 6 months was observed in 5 patients, from 6 months to 1 year - in 7, from 1 year to 2 years - in 7, from 2 to 5 - in 13 and from 5 to 12 years - in 4.

 The lesions in the vast majority of patients (in 28 of 36) were localized on the leaf, in one they were multiple. The size of foci from 0.5 to 1 cm was noted in 30 and from 1 to 1.5 cm in diameter - in 6 patients.

 The lesions were scaly patches of dark color with clear boundaries and a slightly rough surface or plaques of round, oval or irregular shape. Plaques were often covered with a grayish dense crust or papillary outgrowths.

 There were 2 patients with Boven's disease (a man of 67 years and a woman of 47 years). The size of the lesion in a man was 2 x 3 cm, the duration of the disease was 2 years, localization was the left iliac region; the size of the lesion in a woman was 1.5 x 2 cm, the duration of the disease was 11 years, localization in the interscapular region.

 In these patients, lesions were yellowish-red plaques of irregular shape with clear boundaries. The edges of the plaques were slightly raised. On the surface of the foci there were warty growths, small-plate peeling, and bloody crusts.

Treatment technique
Patients were treated on an outpatient basis with applications of 30% ointment of glyciphone composition: glyciphone 30.0, anhydrous lanolin 60.0, petrolatum 10.0. Before applying the ointment, the skin in the area and around it was wiped with a cotton ball soaked in alcohol. Around the hearth, 0.5 cm backward from its edge, zinc paste was applied. Then, glyciphon ointment was applied to the lesion with a 0.5 cm capture of the surrounding skin and a napkin of 4 layers of gauze was applied. A piece of wax paper was inserted between the two layers of the napkin so that the ointment did not soak the upper layers of the napkin. The napkin was fixed with a sticky patch. Glyciphon ointment applications were made 1 time per day. For each dressing, necrotic tissue was removed from the surface of the lesion.

 For senile keratoses, 5 to 10 applications were made (depending on the size of the lesion). Subsequently, aseptic dressings were applied to the lesion sites until complete healing. Defects formed at the site of foci during treatment healed within 7-10 days.

Treatment results
All patients 38 achieved persistent clinical recovery. Duration of observation from 2 to 5 years. In the treatment of patients with senile keratoses and Boven’s disease, no side effects were detected. The general condition of the patients was not impaired. Peripheral blood counts did not change.

Conclusion
Glyciphon ointment 30% is an effective treatment for skin precancerosis.

 Thus, the above results demonstrate the high efficiency of glyciphone ointment in the treatment of precancerosis and skin cancer.

 Based on the results of clinical trials, the pharmacological committee recommended to allow medical use in adults and children as a local antitumor agent for skin cancer, basal cell carcinomas, keratoses, etc. and the industrial release of the drug glyciphone in the form of 30% ointment (protocol N 7 from 04/11/1990. )

 After the development of normative and technical documentation, the Ministry of Health and Medical Industry of the Russian Federation by order No. 4 of 02.24.94 approved the Temporary Pharmacopoeial Articles VFS 42-2277-93 on glyciphone - substance and VFS 42-2276-93 on glyciphon ointment and approved the use in medicine and industrial production of glyciphone and its ointment.

 The active substance of the claimed medicinal product (glyciphon-substance) is synthesized by known methods, for example, by reacting methylphosphonic acid dichloride with glycidyl in the presence of caustic potassium as an acceptor of released hydrogen chloride and in methylene chloride medium. KCl is filtered from the reaction mass, the filtrate is evaporated, the target product is isolated from the residue after evaporation by distillation in vacuo.

The dosage form of glyciphone in vitro is prepared as follows. The ointment base (anhydrous lanolin, petroleum jelly, emulsion waxes) is melted in a porcelain cup at a temperature of 60-70 ^o C, hot filtered through a nylon cloth on a hot filter funnel in a laboratory ointment boiler. Glyciphon-substance is added to the base and mixed using an agitator with inclined blades for 1 hour at an agitator speed of 100 rpm and a temperature of 38 ^o C. Then, for 15 minutes, the ointment is manually mixed with a spatula. Ready ointment is Packed. The ointment is yellow, yellow-brown or greenish-brown in color, with a peculiar smell.

 Industrial applicability. The inventive drug is used in medicine. The release of the first pilot series of 30% glycyphonic ointment is planned at the Tatkhimpharmpreparat KPFO. (Kazan).

Claims (1)

1. A drug for the treatment of cancer and precancerous skin diseases, including an active substance with glycidyl groups and a pharmacological diluent, characterized in that it contains methylphosphonic acid diglycidyl ether of the general formula

and as a diluent - lanolin, petrolatum in the following ratio of components, wt.%:
Methylphosphonic acid diglycidyl ether - 5 - 50
Anhydrous lanolin - 50 - 94
Vaseline - No more than 45
2. The drug according to claim 1, characterized in that it further comprises emulsion waxes in an amount of not more than 10%.

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