WO2003022260A1

WO2003022260A1 - Use of methotrexate and l-arginine for the preparation of a medicament for treatment of uterine myoma

Info

Publication number: WO2003022260A1
Application number: PCT/YU2002/000017
Authority: WO
Inventors: Slobodan Arsenijevic; Zoran Matovic
Original assignee: Individual
Current assignee: Individual
Priority date: 2001-09-12
Filing date: 2002-09-10
Publication date: 2003-03-20
Anticipated expiration: 2004-03-12
Also published as: WO2003022260B1

Abstract

The invention illustrates a drug for treatment of myoma of the uterus that enables preservation of biological and sexual properties of the women and does not provoke contraindications accompanying other known methods of treatment. The invention is represented by a mixture of two substances methotrexate and L-arginine. The drug for treatment of myoma of the uterus is composed of 5 g methotrexate and 10 g L-arginine with 985 cm3 water. Combination of two substances in aqueous solution yields a homogenous mixture where each of the components preserves its own chemical identity. The drug is applied on the eighth day of the follicular phase of the menstrual cycle of patients with myoma of the uterus. The patient is placed into lythotomy position, the vaginal speculum is placed and the uterine cervix is pulled with roller forceps so that the external uterine cervix is exposed. The package is open, the puncture needle iS inserted into the cervical channel and is directed to the myometrium towards the side at which myoma is located. The needle is pushed through the endometrium to the depth of 1 cm. It is necessary to be sure to inject the drug at 1 cm depth to avoid possible penetration through the uterine wall. We aspirate to check that the needle is not inside a blood vessel, and after that inject the drug slowly. The syringe and needle are discarded after use. The mixture is applied in three consecutive menstrual cycles. The method is simple, feasible on out-patient basis by practicing gynecologist; no ultrasonografic guidance is needed; transvaginal and transcervical approach is used. The mixture induces the process of apoptosis (programmed cell death) so that no necrosis, degeneration or inflammation are provoked if all antisepsis precautionary measures are applied. Dramatic reduction of both loss of blood during menstruation in women with myoma and the volume of myoma are evidenced. Since the dose applied is very low, toxicity is negligible. Application of this mixture gradually leads to menopause without disturbance of the hypothalamo-pituitary-gonadal axis. Application of this mixture does not provoke early or late complication specific for other medical approaches listed in the background of the invention. After completion of therapy with this medication, the volume of myoma is reduced by 20-30% and no surgical removal is required. After such therapy myoma does not grow any longer. After the therapy menstrual bleedings become normal in volume.

Description

MEDICATION FOR TREATMENT OF UTERINE MYOMA

Description of invention

Field of invention

The invention belongs to the field of pharmacology and medicine i.e. gynecology, since it is a drug for conservative treatment of uterine myoma.

According to International Patent Classification (IPC) the invention is classified as: A 61 K 31/195.

Technical problem

The technical problem solved by the invention is the following: How to reduce the volume of uterine myoma, reduce the bleeding to a normal level without surgical intervention, without implementation of the existing therapeutic methods that are associated with numerous contraindications and, at the same time preserve all biological and sexual features of the female.

Myomas are the most commonly encountered benign tumors of the uterus. As many as 30-50% perimenopausal women suffer from myoma. Up to about 10 years ago the therapy was surgical and the problems of these women was managed by total hysterectomy with bilateral adnexetomy leading the patients to early menopause immediately.

Nowadays the following alternatives are available for the treatment of uterine myoma: progestagen, danazol, GnRH, embolization of myoma blood vessels by polyvinyl- alcohol. All these methods are associated with numerous adverse effects (disturbance of hypothalamo-pituitary-gonadal axis, abrupt onset of early menopause, necessitated anesthesia, inflammation, etc

This invention solves (cures) the uterine myoma disease without adverse effects that commonly accompany the current therapeutic methods by decreasing hemorrhage (which is the main symptom of uterine myoma), decreases the myoma volume and the patient, following the prescribed procedure, enters her menopause normally, without any surgical intervention.

Background of invention In reference literature related to patents and other no treatment has yet been described to offer therapy of myoma of the uterus without contraindications associated with the known therapeutic methods that would make it possible for the patient to preserve all her biological and sexual features.

Current understanding of sickness and health as well as increasing need for prolongation of active sex life imply preservation of the internal sex organs. Conservative treatment of myoma of the uterus, thus, has become both challenge and need.

The therapy may be non-specific, when use of medication relieves the symptoms that result from the presence of myoma of the uterus and specific that in addition to relief of the symptoms also reduce the volume of the myoma of the uterus, uterine hemorrhage and improve hematological status of the patient.

In principle, pharmacotherapy of the myoma of the uterus can be used as monotherapy of supplementary. Medical therapy not only solves the problems of many patients, but frequently helps surgical therapy be less radical and safer for the patient.

The earliest conservative method for treatment of myoma of the uterus implied use of progestagen, whose role is to stop the proliferative changes in the endometrium, i.e. indirectly to stop abundant and prolonged uterine bleeding in women suffering from myoma of the uterus. This method is associated with ovarian cyst as the adverse effect, while the method itself only reduces the volume of bleeding while the myoma continues to grow.

Another method that is used with more or less success in order to reduce or stop heavy menstrual bleeding is use of danazol, antigonadotropin that reversibly influences the impact of gonadotropic hormones. Continuous application of these effectively reduces the volume of menstrual bleeding and brings the patient to the state of secondary amenorrhea, but the adverse effects are still numerous and dose related: hot flashes, heavy sweating, occurrence of acnae, hirsutism, weight gain and seborrhea.

Gn H analogues have been used for ten years now, and their continuous administrations irreversibly suppresses gonadal function leading to a condition similar to that hypogonadotropic hypogonadism. Long-term administration leads to discontinuation of estrogen secretion, resulting in discontinuation of menstrual bleeding, reduction and regression of myoma and improvement of the hematological status. Adverse effects include occurrence and signs of post-menopause: hot flashes and dry vagina encountered in over 90%) patients, while insomnia and depression are also quite common. Early occurrence of osteoporosis is the most serious adverse effect. Frequent adverse effects and associated risks are limiting factors of this therapy, while application of GnRH analogues disturbs the natural cycle of hormone secretion.

Vascular sclerozing of myoma of the uterus by alcohol is an effective non- surgical method aimed at selective sclerosing of the blood vessels that supply the myoma. Antibody sclerosis reduces supply to the myocytes, resulting in their degeneration with resultant necrosis and degeneration. The method is aggressive, associated with risk and may result in immediate complications.

Embolization of blood vessels is a method similar to the previously described one, where polyvinyl alcohol is injected into the myomal vessels to block them, preventing blood supply to the myoma vessels which may lead in aseptic necrosis and degeneration accompanied with sharp pain immediately after the procedure and occurrence of high fever.

The aim of the invention to reduce the volume of the myoma of the uterus, reduce the bleeding to a normal level, and avoid all the above complications and preserve biological and sexual features of the woman.

Summary of the invention

The invention is represented by a mixture of two substances.

The first substance is methotrexate: 4-amino-N-lO-methyl-pteroilglutamic acid; CASRN: 59-05-2; C₂₀H₂₂N₈O₅; Molecular mass = 454.4444; insoluble in water (<0.1 g/100 mL at 19 °C); yellow to dark orange crystal powder; hygroscopic, sensitive to light. Methotrexate is a drug used for treatment of some malignant diseases, severe forms of rheumatoid arthritis and intact extrauterine pregnancy, hydatiform mole and choriocarcinoma.

The second substance is L-arginin; 2-amino-5-guanidino- pentanic acid; CASRN: 74-79-3; C₆H_MN₄O₂; Molecular mass = 174.2022; basic amino acid; essential amino acid; white powder.

Combination of two substances in aqueous solution yields a homogenous mixture where each of the components preserves its own chemical identity.

Local application of aqueous solution of the mixture of these two substances subendometrially into the myoma on the eighth day of the follicular phase of the menstrual cycle bleeding results in reduction of the myomal volume. The mixture is applied in three consecutive menstrual cycles. The method is simple, feasible on out-patient basis by practicing gynecologist; no untrasonografic guidance is needed; transvaginal and transcervical approach is used. The mixture induces the process of apoptosis (programmed cell death) so that no necrosis, degeneration or inflammation are provoked if all antisepsis precautionary measures are applied. Dramatic reduction of both loss of blood during menstruation in women with myoma and the volume of myoma are evidenced. Application of this mixture gradually leads to menopause without disturbance of the hypothalamo- pituitary-gonadal axis.

The drug is most effective in treatment of myoma of the uterus with diameter up to 10 cm.

After completion of therapy with this medication, the volume of myoma is reduced by 20-30% and no surgical removal is required. After such therapy myoma does not grow any longer. After the therapy menstrual bleedings become normal in volume. The patient continues her normal life with preserved biological and sexual features until normal menopause when menstrual bleeding is discontinued and myoma of the uterus stops growing. The aim of the therapy is to bridge over the period to menopause with preserved biological and sexual features of the woman, i.e. without any adverse effects on her health.

Application of the drug does not induce any direct or indirect late complications specific for other medical treatments cited above.

Summary of the illustrations Figure 1. illustrates molecular structures of methotrexate and L-arginine. Figure 2. gives the illustration of molecular appearance of the homogenous mixture of methotrexate and L-arginine.

Detailed description of the invention This pharmaceutical composition contains: - 5 g methotrexate (CASRN: 59-05-2)

10 g L-arginine (CASRN: 74-79-3) 985 cm³ water.

The first substance is methotrexate: 4-amino-N-lO-methyl-pteroilglutamic acid; CASRN: 59-05-2; C₂₀H₂₂N₈O₅; Molecular mass = 454.4444; insoluble in water (<0.1 g/100 mL at 19 °C); yellow to dark orange crystal powder; hygroscopic sensitive to light. Methotrexate is a drug used for treatment of some malignant diseases, severe forms of rheumatoid arthritis and intact extrauterine pregnancy, hydatiform mole and choriocarcinoma.

The second substance is L-arginin; 2-amino-5-guanidino- pentanic acid; CASRN: 74-79-3; C₆Hι₄N₄O₂; Molecular mass = 174.2022; basic amino acid; essential amino acid; white powder.

Figure 1. illustrates molecular structures of methotrexate i L-arginine. Dissolution of methotrexate and L-arginine in water results in formation of homogenous mixture without any chemical reactions that might change the chemical structure of either of the components. However, some secondary interactions do take place in the homogenous mixture (Figure 2.) caused by acid-base reaction of methotrexate and L-arginine as well as the occurrence of hydrogen bond. L-Arginine as a very alkaline amino acid deprotonizes the carboxyl groups of methotrexate. The hydrogen bond between the methotrexate and L- arginine molecules is realized primarily between the oxygen atoms of the carboxyl groups of methotrexate and nitrogen atoms of L-arginine, as well as oxygen atom of the L-arginine carboxyl group and nitrogen atoms of methotrexate. Molecules of water are included in the network of the above hydrogen bonds.

Use the analytical balance (precision to the fifth decimal number) to weigh 10 g L-arginine and transfer quantitatively to a normal laboratory vessel of 1 dm . After that 600 cm³ of re-distilled water is added to the vessel. Close the vessel and shake slowly until L-arginine is completely dissolved. Completely colorless solution is obtained. Use the analytical balance (precision to the fifth decimal number) to weigh 5 g methotrexate and transfer quantitatively to the laboratory vessel containing L-arginin solution. Shake the vessel mildly until methotrexate is completely dissolved. Fill to the volume mark with redistilled water. Place the solution into standard autoclave for processing and store in refrigerator at 4 to 8 °C.

The single dose is 2 cm³ of the drug solution in a 2 cm³ syringe with a 20 G puncture needle. The set is sterilized.

Mode of application The drug is applied on the eighth day of the follicular phase of the menstrual cycle in patients with myoma of the uterus. The single dose is 2 cm³ of the drug solution in a 2 cm syringe with a 20 G puncture needle. The set is sterilized.

The patient is placed into lythotomy position, the vaginal speculum is. placed and the uterine cervix is pulled with roller forceps so that the external uterine cervix is exposed. The package is open, the puncture needle is inserted into the cervical channel and is directed to the myometrium towards the side at which myoma is located. The needle is pushed through the endometrium to the depth of 1 cm. It is necessary to be sure to inject the drug at 1 cm depth to avoid possible penetration through the uterine wall. We aspirate to check that the needle is not inside a blood vessel, and after that inject the drug slowly. The syringe and needle are discarded after use. The procedure is repeated in three consecutive menstrual cycles.

After completion of therapy with this medication, the volume of myoma is reduced by 20-30%) and no surgical removal is required. After such therapy myoma does not grow any longer. After the therapy menstrual bleedings become normal in volume.

Claims

PATENT CLAIMS

1. Product, aqueous solution of mixture of methotrexate and L-arginine wherein 5 g methotrexate and 10 g L-arginineare are mixed in 985 cm of water.

2. Process of aqueous solution of the mixture production pursuant to this patent claim 1. weherein 10 g L-arginine is weighed on analytical balance and quantitatively transferred to a 1 dm vessel where 600 cm water is added, mildly shaken until L-arginin is completely dissolved. After that 5 g methotrexate is added, the vessel is mildly shaken until all methotrexate is completely dissolved, the remaining water is added, and the obtained solution is placed in a standard autoclave for processing.

3. The aqueous solution of the mixture is used, pursuant to patent claim 1 for treatment of myoma of the uterus.