EA013430B1 - Testosterone gels comprising propylene glycol as penetration enhancer - Google Patents

Abstract

The present invention relates to testosterone-containing pharmaceutical compositions and gels, and to methods of using the same.

Description

FIELD OF THE INVENTION

The present invention relates to pharmaceutical compositions and gels containing testosterone, and methods for their use.

State of the art

Testosterone in men.

Testosterone, the main circulating androgen in men, is synthesized from cholesterol. Approximately 500 million Leydig cells in the testes secrete more than 95% of 6-7 mg of testosterone produced daily. Two hormones produced by the pituitary gland, luteinizing hormone (LH) and follicle-stimulating hormone (EH), are necessary for the development and maintenance of testis function and for the negatively regulated formation of testosterone. Circulating testosterone is metabolized to various 17-ketosteroids in two different ways. Testosterone can be metabolized to dihydrotestosterone (INT) by the enzyme 5-alpha reductase or to estradiol (E2) by the aromatase enzyme complex. About 90% of testosterone circulating in the blood is protein bound. In men, approximately 40% of the binding is to high affinity sex hormone binding globulin (8HBC). The remaining 60% are weakly bound to albumin. So, there is a lot of data on testosterone determination provided by clinical laboratories.

The table in the University of California Medical Center Los Angeles Medical Center (ICA-NagLog Mob1e1 Sep1g) below summarizes hormone concentrations in normal adult males.

Hormone levels in normal men

Testosterone (total)

Free testosterone

ENT

ΫΗΤ / Τ ϋΗΤ + Τ ratio

8NVS

G8N

Bh

E2

The range of normal hormone concentrations is from 298 to 1043 ng / dl from 3.5 to 17.9 ng / dl from 31 to 193 ng / dl from 0.052 to 0.33 from 372 to 1349 ng / dl from 10.8 to 46.6 nmol / L from 1.0 to 6.9 mIU / ml from 1.0 to 8.1 mIU / ml from 17.1 to 46.1 pg / ml

According to literature there is a significant difference in the half-life of testosterone, ranging from 10 to 100 minutes. Researchers, however, agree that circulating testosterone in a normal young man differs over the course of a day. The maximum level falls on the period from 6:00 to 8:00 in the morning and then decreases during the day. Typical profiles for the maximum testosterone level are 720 ng / dl and for the minimum level - 430 ng / dl. However, the physiological significance of this diurnal cycle, if one exists, is not known.

Testosterone in women.

The excretion of androgenic steroids into the urine of adult women was demonstrated more than 50 years ago. Since that time, physiologists and clinicians have studied the sources and biological functions of testosterone and other endogenous androgenic hormones in women. It is now known that androgens in women are secreted by both the ovaries and adrenal glands. Each of these sources contributes approximately 50% (directly or with the participation of predecessors) to approximately 300 μg of testosterone produced daily in healthy women of reproductive age (“cycling women”). Despite the fact that the adverse effects of androgen overproduction found in polycystic ovary syndrome and some tumors producing androgens have been well established, the normal physiological effects of androgens in women are much less understood. As suggested by animal studies, men's physiology, and symptoms in women with insufficient androgen production, the main physiological effects of androgens in normal women include, but are not limited to, anabolic effects on muscles, skin, hair, and bones; stimulating effects on erythropoiesis; modulating effect on immunity and physiological effects on mood, health and sexual function.

In addition, endogenous androgens are important for the development of pubic hair and it is believed that they modulate the action of estrogens and progestins on a variety of reproductive target tissues. Androgens are believed to play an important role in modulating the secretory function of the lacrimal gland.

50% of circulating testosterone is formed as a result of direct secretion of the ovaries in the tecal cells under the control of luteinizing hormone. The other half is the result of peripheral conversion of the adrenal androgenic precursors of dehydroepiandrosterone, androstenedione, and dehydroepiandrosterone sulfate. Testosterone can also be converted to dihydrotest

- 1 013430 sterone or estradiol. Thus, testosterone is both a hormone and a pre-hormone.

About 98% of the testosterone circulating in the blood is protein bound. In women, approximately 66% of binding is accounted for by high affinity sex hormone binding globulin (8NBO). The remaining 34% are weakly bound to albumin. So, there is a lot of data on testosterone determination provided by clinical laboratories. The location of steroids that are most strongly associated with high-affinity sex hormone-binding globulin according to their affinity is: dihydrotestosterone> testosterone> androstenedione> estradiol> estrone. High-affinity sex hormone binding globulin weakly binds dihydrotestosterone, but not dihydrotestosterone sulfate. The table below shows hormone levels in normal women before menopause.

Hormone levels in normal women before menopause

Hormone Average +/- 8B Middle Range Testosterone (nmol / l) 1.2 +/- 0.69 level 0.98 0.4-2.7 Free Testosterone (pmol / L) 12.80 +/- 5.59 12.53 4.1-24.2 % free testosterone of total 1.4 +/- 1.1 1,1 0.4-6.3 testosterone Luteinizing hormone (MED / L) 7.2 +/- 3.3 6.7 3.0-18.7 Follicle-stimulating hormone 4.7 +/- 3.6 4.2 1,5-21,4 (MED / L) High affinity globulin 66.1 +/- 22.7 71.0 17.8-114.0 sex hormone binding (nmol / L)

Compared with deficiency states for other hormones, testosterone deficiency in women was practically ignored as a clinical manifestation. However, there are well-defined populations of subjects in which the production of androgens is clearly inadequate and in which the associated symptoms are described. These populations include, for example, young women with ovarian / uterus removed, postmenopausal women undergoing estrogen replacement therapy, women taking oral contraceptives, women with adrenal dysfunction, women with adrenal suppression caused by corticosteroids, and women positive for the AIDS virus .

Since it has been shown that increasing testosterone concentrations changes sexual activity or libido, the researchers studied ways to deliver testosterone to men as well as women. These methods include intramuscular injection, oral replenishment, ball implants, and transdermal patches.

These testosterone delivery methods, however, have one or more disadvantages. For example, subcutaneous ball implants and ester injections are painful and require surgical procedures and / or doctor visits. Moreover, hypogonadal male implant therapy includes the risk of expulsion (8.5%), bleeding (2.3%) or infection (0.6%). Many of these methods, such as the use of oral / sublingual / buccal preparations, suffer from an undesirable pharmacokinetic profile created by superphysiological concentrations of testosterone, which then return to baseline. Transdermal patches provide less than optimal pharmacokinetic characteristics, constrain many subjects, and are associated with significant skin irritation. In addition, the patches do not provide flexible dosing, may be visually unattractive, and may tend to fall off, especially during intense physical exercise. Oral administration creates inappropriate testosterone levels and drug uptake unpredictable among various subjects. Moreover, since the liver metabolizes the drug, there is a risk of hepatotoxicity, not to mention the presystemic metabolism.

Recently, a one percent testosterone gel for use in men has been approved, which provides flexible dosing with minimal skin irritation. This gel is available in the United States under the trademark ApbgoOe1® (Sytel Ryagtaseijsa S, 1ps., BeegjeM, III., A 8o1uauRNagtaseijsa18, 1ps. So., Mapeye, OA 30062).

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Composition LpbgoSe1®

ΑηάτοΟεΙ® Composition Amount (weight / mass) per 100 g of gel Testosterone 1.0 g Sagoro 1980 0.90 g Isopropyl myristate 0.50 g Ο, ΙΝΝηΟΗ 4.72 g Ethanol (95% vol / vol) 72.5 g * Purified Water (DZ!) 100 g

Corresponds to 67 g of absolute ethanol.

The present invention provides pharmaceutical compositions and gels containing testosterone, and methods for their use. In more detail, the present invention provides hydroalcoholic / alcoholic pharmaceutical compositions and gels containing testosterone suitable for topical and transdermal application, said compositions containing propylene glycol. In addition to achieving very effective testosterone delivery, the compositions of the invention are aesthetically attractive. Propylene glycol has been shown to be a very powerful permeability enhancer in the composition of the invention, which was used as a single permeability enhancer or in conjunction with an additional permeation enhancer (s), for example in conjunction with isopropyl myristate.

The present invention relates to a pharmaceutical composition comprising testosterone, at least one C ₂ -C ₆ alcohol, at least one thickener, at least one penetration enhancer and water.

The specified pharmaceutical composition can be prepared in various forms, for example in the form of a gel, solution, cream, lotion, spray solution, ointment, aerosol. Preferably, said pharmaceutical composition is a gel.

The term “testosterone” as used herein refers not only to testosterone per se, but also to its enantiomers, isomers, tautomers, salts, chelates, complex alcohols, amides, derivatives and prodrugs and their precursors, for example ΌΗΤ (dihydrosterone), 17-methyltestosterone, 3-cyclopentyl ester of 17 [alpha] -methyltestosterone-enol, testosterone enanthate, testosterone cypionate, testosterone undecanoate, testosterone cyclodextrin, testosterone bicyclate. In accordance with the present invention, testosterone may be of natural origin or the result of semisynthetic or synthetic processes.

C ₂ -C ₆ alcohols are known in the art. Such alcohols include ethanol, propanol, isopropanol (propan-2-ol), n-propanol (propan-1-ol), butanol, butan-1-ol, butan-2-ol, tert-butanol, pentanols, hexanols. Ethanol is preferable, as it greatly facilitates the transdermal transfer of testosterone due to its rapid evaporation upon contact with the skin.

Thickeners are known in the art. The term “thickeners” generally refers to a compound, possibly of a polymer nature, having the ability to gel into contact with a specific solvent, for example water. Thickeners make it possible to increase the viscosity of the pharmaceutical compositions in accordance with the present invention, but can also act as solubilizing agents. Examples of thickeners include anionic polymers such as acrylic acid-based polymers (including polyacrylic acid polymers such as SLKVOROB® (B.R. carbomers or polymers based on acrylic acid, for example Cagoro1® 980 or 940, 981 or 941, 1382 or 1382, 5984, 2984, 934 or 934P (Cagoro1® are usually acrylic acid polymers crosslinked with allyl sucrose or allylpentaerythritol), Ret1 ® or TK2® (ϋΐΐ ^ ζ, 8in1a1ep SK, e! e.); cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (SMS), etc .; poloxamers or copolymers of polyethylene polypropylene1 such as 127, poloxamines and other thickeners, such as chitosan, dextran, pectins and natural gums, all of these thickeners, alone or as a mixture, can be used in the pharmaceutical composition of the present invention. The above thickeners can be selected, taking into account the pH of the composition in accordance with the present invention and the desired viscosity.

In the context of the present invention, hydroxypropyl cellulose, Cactoro® 980 and Bilco1® are preferred.

- 3 013430

Permeation enhancers are also known in the art. A “permeation enhancer” is usually a substance for which it is known that it enhances the delivery of a drug or active ingredient through the skin. These compounds can also be attributed to permeability accelerators, activating additives and absorption promoters. This class of substances includes compounds with a variety of mechanisms of action, including mechanisms whose function is to improve the solubility and diffusion of the drug, and mechanisms that improve transdermal absorption by modifying the ability of the stratum corneum to retain moisture, softening the skin, improving skin permeability, acting as helpers permeability or compounds loosening hair follicles, or temporarily changing the condition of the skin, such as a boundary layer.

Unless expressly stated, percentages (%) refer to the mass amounts of the substance relative to the total weight of the compositions.

In accordance with one aspect of the specified pharmaceutical composition comprises from 0.5 to 5.0% (wt./wt.), Preferably from 0.5 to 2.5% (wt./wt.), More preferably from 0.6 up to 2.0% (w / w), even more preferably from 0.7 to 1.5% (w / w), even more preferably from 0.75 to 1.25% (w / w) .), even more preferably from 0.8 to 1.2% (wt./wt.) and even more preferably from 0.9 to 1.1% (wt./wt.), most preferably about 1% (wt. ./mass.) testosterone.

In accordance with another aspect of the present invention, said pharmaceutical composition comprises from 0.5 to 5.0% (w / w), preferably from 0.5 to 2.5% (w / w), more preferably from 0 75 to 2.25% (w / w), even more preferably from 0.9 to 2.0% (w / w), even more preferably from 1.0 to 1.8% (w / w) / wt.), even more preferably from 1.25 to 1.75% (wt./wt.), even more preferably from 1.3 to 1.6% (wt./wt.), most preferably about 1, 5% (w / w) testosterone.

In accordance with another aspect of the present invention, said pharmaceutical composition comprises from 40.0 to 75.0% (w / w), preferably from 40.0 to 70.0% (w / w), more preferably from 45 0 to 65.0% (w / w), even more preferably from 50.0 to 60.0% (w / w), even more preferably from 52.0 to 58.0% (w / w) / wt.), even more preferably from 53.0 to 57.0% (wt./wt.), even more preferably from 54.0 to 56.0% (wt./wt.), most preferably about 56, 0% (w / w) of at least one C ₂ -C ₆ alcohol.

In accordance with another aspect of the present invention, said pharmaceutical composition comprises from 0.1 to 5.0% (w / w), preferably from 0.15 to 4.5% (w / w), more preferably from 0 2 to 4.0% (w / w), even more preferably 0.25 to 3.5% (w / w), even more preferably 0.3 to 3.0% (w / w) / wt.), even more preferably from 0.4 to 2.5% (wt./wt.), even more preferably from 0.5 to 2.0% (wt./wt.), most preferably from about 0 5 to 1.0% (w / w) of at least one thickener.

In accordance with another aspect of the present invention, said thickener is selected from the group consisting of acrylic acid-based polymers, including cellulosic polymers, including cellulose ethers and derivatives (cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose (HPMC), carboxymethyl methyl cellulose (HPMC) ) etc.); carbomers, such as Cagoro1® polymers, for example Cagoro1®980 or 940, 981 or 941, 1382 or 1382, 5984, 2984, 934 or 934P, Retiep TK1® or TK2®, yigskh. 8up1yaya1 SK, etc .; poloxamines, poloxamers or copolymers of polyethylene-polypropylene, such as Li1go1® grades 68 or 127, poloxamines and other thickeners such as chitosan, dextran, pectins and natural gums, and mixtures thereof.

In accordance with another aspect of the present invention, said pharmaceutical composition comprises from 0.1 to 5.0% (w / w), preferably from 0.15 to 4.5% (w / w), more preferably from 0 2 to 4.0% (w / w), even more preferably 0.3 to 3.5% (w / w), even more preferably 0.4 to 3.0% (w / w) / wt.), even more preferably from 0.5 to 2.5% (wt./wt.), more preferably from 0.5 to 2.0% (wt./wt.) of propylene glycol. Advantageously, in accordance with the present invention, for the content of testosterone and compositions of the invention, it has been found that propylene glycol is a very effective and compatible penetration enhancer. In addition, the use of propylene glycol in accordance with the present invention leads to pharmaceutical compositions having a very attractive appearance. The pharmaceutical compositions of the invention are very transparent, visually attractive and have a very pleasant texture that improves the use of the compositions by the patient. In addition, the pharmaceutical compositions of the invention require lower alcohol concentrations. Moreover, the compositions of the invention are very stable, and it has been proven that propylene glycol is compatible with standard neutralizing agents, for example triethanolamine. Compositions in accordance with the invention may include several permeation enhancers.

In accordance with the present invention, said composition optionally further comprises water.

- 4 013430

Said pharmaceutical composition is suitable for controlled transdermal delivery of testosterone. Thus, the specified pharmaceutical composition can be taken by the patient and does not require the presence of a doctor at the time of administration. Also, the introduction does not require surgical procedures or injections.

Advantageously, in accordance with the present invention, propylene glycol acts as a very effective penetration enhancer for testosterone, in particular in combination with a dosage of testosterone and a thickener (thickeners) of the present composition.

In another aspect, the pharmaceutical composition of the present invention further comprises a base. Advantageously, said base is pharmaceutically acceptable, the base is preferably selected from the group consisting of triethanolamine, sodium hydroxide, ammonium hydroxide, potassium hydroxide, arginine, aminomethylpropanol or tromethamine and mixtures thereof. If the pH of said pharmaceutical composition is not optimized for transdermal administration, for example, if said thickener includes at least an acrylic acid polymer, said base helps to neutralize said pharmaceutical composition for topical application to human skin. In addition, this base (neutralizing substance) promotes optimal swelling of the polymer chains during charge neutralization and the formation of polymer salts. In particular, if said thickener comprises an acrylic acid polymer, said base preferably includes triethanolamine. It also helps to achieve optimal viscosity in the pharmaceutical composition in accordance with the present invention. Specialists in this field of technology know how to choose a suitable amount of the specified base in the composition, especially regarding the nature of the specified thickener and the alcohol content in the composition, in order to achieve the desired final pH value in the composition. For example, in the presence of carbomers and / or if the alcohol content is high, tromethamine and / or ΝαΝ can be used as the base in amounts selected in such a way as to achieve the desired final pH in the composition. Alternatively, in one embodiment, the pharmaceutical composition of the present invention comprises from 0.1 to 5.0% (w / w), preferably from 0.15 to 4.5% (w / w), more preferably 0.2 to 4.0% (w / w), even more preferably 0.25 to 3.5% (w / w), even more preferably 0.3 to 3.0% (w / w), even more preferably from 0.4 to 2.5% (w / w), even more preferably from 0.5 to 2.0% (w / w), most preferably from 0.5 to 1.0% (w / w) triethanolamine.

Preferably, the pH of the pharmaceutical composition in accordance with the present invention would be between 2 and 9, preferably between 3 and 8, even more preferably between 3 and 7. Said pharmaceutical composition of the present invention further comprises isopropyl myristate. In accordance with the present invention, it was unexpectedly shown that the addition of isopropyl myristate leads to a significant improvement in results when used together with propylene glycol as an additional permeability enhancer. Preferably, said pharmaceutical composition comprises from 0.05 to 5.0% (w / w), more preferably from 0.1 to 4.0% (w / w), more preferably from 0.15 to 3, 0% (w / w), even more preferably 0.25 to 2.5% (w / w), even more preferably 0.25 to 2.0% (w / w), even more preferably 0.25 to 1.5% (w / w), even more preferably 0.25 to 1.0% (w / w), most preferably 0.5 to 0.75 % (w / w) isopropyl myristate.

Thus, in one embodiment, the present invention provides a pharmaceutical composition comprising:

from 0.5 to 5.0% (w / w) testosterone;

from 40.0 to 75.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 5.0% (w / w) of at least one thickener;

from 0.1 to 5.0% (w / w) propylene glycol;

from 0.1 to 4.0% (w / w) isopropyl myristate; optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: from 0.75 to 1.25% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 4.0% (w / w) of at least one thickener;

0.2 to 4.0% (w / w) propylene glycol;

from 0.15 to 2.0% (w / w) isopropyl myristate; optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: from 0.8 to 1.2% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 3.0% (w / w) of at least one thickener;

from 0.3 to 3.0% (w / w) propylene glycol;

from 0.35 to 1.5% (w / w) isopropyl myristate;

- 5 013430 optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: from 0.9 to 1.1% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 2.5% (w / w) of at least one thickener;

from 0.4 to 2.5% (w / w) propylene glycol;

from 0.4 to 1.0% (w / w) isopropyl myristate;

optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: about 1.0% (w / w) testosterone;

from 50.0 to 70.0% (w / w) to at least one C ₂ -C ₆ alcohol;

from 0.1 to 2.5% (w / w) of at least one thickener;

from 0.4 to 2.5% (w / w) propylene glycol;

about 0.5% (w / w) isopropyl myristate;

optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: from 1.0 to 2.0% (w / w) testosterone;

from 50.0 to 75.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 5.0% (w / w) of at least one thickener;

from 0.1 to 5.0% (w / w) propylene glycol;

from 0.1 to 4.0% (w / w) isopropyl myristate;

optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: 1.25 to 1.75% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 4.0% (w / w) of at least one thickener;

0.2 to 4.0% (w / w) propylene glycol;

from 0.15 to 2.0% (w / w) isopropyl myristate;

optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: from 1.4 to 1.6% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 3.0% (w / w) of at least one thickener;

from 0.3 to 3.0% (w / w) propylene glycol;

from 0.35 to 1.5% (w / w) isopropyl myristate;

optionally water.

In another aspect, the present invention provides a pharmaceutical composition comprising: about 1.5% (w / w) testosterone;

from 50.0 to 70.0% (w / w) of at least one C ₂ -C ₆ alcohol;

from 0.1 to 2.5% (w / w) of at least one thickener;

from 0.4 to 2.5% (w / w) propylene glycol;

from 0.4 to 1.0% (w / w) isopropyl myristate;

optionally water.

In addition, the pharmaceutical composition may include conventional pharmaceutical additives, including salt (s), emollient (s), stabilizer (s), antimicrobial agent (s), flavor (s) and / or propellant (propellants). The pharmaceutical composition may also include at least an additional active ingredient, for example another hormone.

The invention also provides a gel useful for transdermal or transdermal delivery, comprising said pharmaceutical composition in accordance with the present invention. Thus, the invention also targets a hydroalcoholic gel containing testosterone.

According to another embodiment, the present invention provides a dosage unit, a unit dosage unit, and a package containing said pharmaceutical composition or said hydroalcoholic gel in several doses. Preferably, such conditioning of said pharmaceutical composition provides the patient with easier administration. Thus, these forms of packaging may reflect a pattern of use, for example daily or weekly use.

In accordance with another embodiment, a dispenser is provided, for example, a device with a hand pump or valve, comprising said pharmaceutical composition or said hydroalcoholic gel. Such dispensers allow for flexible dosage taking, depending on the amount of applied composition.

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In accordance with another embodiment of the specified packaging or dispensers may be accompanied by notification instructions for their use.

The pharmaceutical compositions, gels, packaging and containers of the present invention are useful in the treatment of testosterone deficiency and in the treatment and / or prevention of conditions and / or disorders associated with testosterone deficiency.

The terms “treating” or “treatment” as used herein refer to any treatment for a condition, disorder, or disease in a mammal associated with a depressive disorder, and include, but are not limited to, preventing a condition, disorder, or disease occurring in a subject that may be predisposed to a condition, disorder or disease, but which has not yet been diagnosed as a patient having a condition, disorder or disease; suppression of a condition, disorder or disease, for example, delayed development of a condition, disorder or disease; relieving a condition, disorder, or disease, for example, stimulating regression of a condition, disorder, or disease; or alleviating a condition caused by a disease or disorder, for example, by stopping the symptoms of the disease or disorder.

The terms "warn" or "warning" in relation to a condition, disorder or disease means the absence of the development of the condition, disorder or disease, if they have not occurred, or the absence of further development of the condition, disorder or disease, if such a development of the condition, disorder or disease has already occurred .

As used herein, the term "testosterone deficiency" refers to lower levels of testosterone in the blood serum of a subject compared to the average level in a healthy subject of the same age. For example, normal women of the reproductive period produce approximately 300 mcg of testosterone per day. Their total serum testosterone levels typically range from about 20 to about 80 ng / dl, averaging around 40 ng / dl. In healthy young women, for example, the average level of free testosterone is usually about 3.6 pg / ml. However, some factors can affect both total serum testosterone levels and serum free testosterone levels. For example, regularly ovulating women have a small but significant increase in plasma testosterone during the middle third of the menstrual cycle. However, the average level of testosterone (1.2 nmol / L or 33 ng / dl) and the average level of free testosterone (12.8 pmol / L or 3.6 pg / ml) during the luteal and follicular phases are not significantly different. In addition, testosterone production is constantly falling after 30 years, so the level of serum testosterone in a 60-year-old woman is only 50% of the level of serum testosterone in a 30-year-old woman. Although the percentage of free testosterone usually does not change with age, an absolute drop in free testosterone was observed. This drop does not occur suddenly during the menopause, but occurs gradually and continuously as a result of an age-related decrease in androgen production by both the adrenal glands and the ovaries. Thus, women begin to experience symptoms associated with menopause in the period immediately preceding menopause. The fall in testosterone followed by menopause is due to a combination of abnormalities in the ovaries, decreased renal secretion and peripheral conversion. In addition, for example, after removal of the ovaries, testosterone concentrations decrease by approximately 50%. The diagnosis of testosterone deficiency is known to the general practitioner practicing in the relevant field of medicine.

The pharmaceutical compositions and gels of the present invention are also useful for treating a variety of physiological and psychological parameters and / or conditions associated with testosterone deficiency in a man or woman. For example, the pharmaceutical compositions and gels of the present invention are useful for enhancing libido and improving sexual efficacy and / or treatment of sexual dysfunction; normalization of hypogonadism;

increasing the mineral density of bones and corresponding markers, increasing the body mass of exhausted people and reducing body weight in obese people, improving muscle mass and efficiency;

normalization of glucose levels; improving symptoms of diabetic retinopathy, as well as reducing the need for insulin in diabetic patients;

treating, preventing, or relieving a cataract attack; treatment, prevention or reduction of manifestations of cardiovascular disease, including the normalization of hypertension; and treatment of obesity;

normalization of cholesterol; normalization of abnormal cardiograms in subjects and treatment, prevention and reduction of vasomotor symptoms;

prevention of debilitating human viral immunodeficiency syndrome;

prevention of osteoporosis, osteopenia, vaginal dryness and thinning of the vaginal wall; relieve symptoms of menopause and hot flashes; treatment of premenstrual syndrome;

treatment, prevention and alleviation of an attack of Alzheimer's disease, dementia;

improve cognitive ability, improve mood and self-esteem; treatment and / or pros

- 7 013430 lactation of depressive disorders; treating, preventing, or reducing cognitive dysfunction; treatment of autoimmune diseases.

The present pharmaceutical compositions and gels can also be used in “combination therapy” in conjunction with another hormone, or steroid, or a pharmaceutical that increases testosterone levels in a subject, or together with estrogen hormone, or another pharmaceutical, such as, for example, an antidepressant .

The invention also relates to a method for producing a pharmaceutical composition, for example in the form of a gel or solution, in accordance with the present invention.

In one aspect, said method comprises the step of dissolving testosterone in a solution of at least one C ₂ -C ₆ alcohol and at least one penetration enhancer (e.g., propylene glycol and / or isopropyl myristate) with stirring.

In another aspect, said method comprises the step of adding water to the resulting mixture with stirring.

If it is desired to obtain a gel, at least one thickening agent, such as Cloro®, is added to the mixture with stirring.

Optionally, a base / neutralizing agent such as triethanolamine is added to the mixture with stirring.

In another aspect, the invention relates to a method for producing a pharmaceutical composition or gel in accordance with the invention, comprising the following steps:

obtaining a mixture containing at least one C ₂ -C ₆ alcohol, at least one penetration enhancer (for example propylene glycol and / or isopropyl myristate) and testosterone;

optionally adding water and mixing;

optionally adding a thickener to the mixture and mixing;

optionally adding a base and stirring again.

In another aspect, the invention also relates to the use of a gel or solution in accordance with the present invention for the manufacture of a medicament for transdermal use in the treatment of a physiological condition associated with androgen / testosterone deficiency.

In another aspect, the present invention relates to a method of treatment.

In one embodiment, said method comprises the step of administering testosterone to a subject in need thereof.

In accordance with one aspect, the pharmaceutical composition or gel of the present invention is applied to healthy skin, for example, to the shoulder or the outside of an arm, thigh, or leg.

To measure and determine the amount of testosterone needed to deliver to a subject in need, serum testosterone concentrations can be measured using standard measurement techniques.

Based on this, qualified personnel will know how to determine the amount of the composition to be taken depending on the exact dosage of testosterone in the specified pharmaceutical composition.

In general, a typical daily dose of a pharmaceutical composition in the form of a gel or solution in accordance with the invention is from 2.5 to 5 g of the composition per day.

The advantages of the invention will become apparent from the following examples, which are presented below as illustrations, but not the basis for limiting the invention.

Those skilled in the art will understand that the present invention may include any number of preferred details described above.

All citations referred to in the text are included in the description by reference in its entirety.

Other embodiments of the present invention, not present here, which are obvious to experts in this field of technology, therefore, also constitute the scope and essence of the present invention. Unless expressly stated, in the practical implementation of the present invention can be applied to the traditional pharmacological and pharmaceutical methods that are in service in this technical field.

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Examples

Example 1

Studies on the transdermal delivery of testosterone in gel formulations were performed on the skin of animals and humans using labeled ( ¹⁴ C) -testosterone and liquid scintillation titration. These studies made it possible to determine the kinetic parameters (amount released after 24 hours and flow). Two aspects were investigated:

physicochemical studies (testosterone solubility and gel viscosity);

percutaneous delivery (the effect of the concentration of testosterone, the nature of the thickener, the presence of permeation enhancers).

Materials and methods

1. Materials.

Reagents

Testosterone (RiKA);

¹⁴ C-Testosterone (ATEGAT).

Thickeners.

Saporo® 934P (Sarb. 934P) (Po1ur1a8ys);

Sagoro1®980 (Sag. 980) (L0! 4419, LarSpez Weszs Wesouzso);

K1ise1®.

Neutralizing agents.

Triethanolamine (1O! 4343, LaBeaSpez Weszs Vouezso).

Permeation enhancers.

Tgap8siapo1 (Oaye Gozze);

oleic acid (OA) (Prgo1aO); propylene glycol (PO) (Oye Gogze);

propylene glycol dipelargonate (ROER) (Oaye Gozze).

Animal skin model.

Male hairless rat, 5 weeks.

Human skin model.

Pieces of biopsy after abdominal plastic surgery, white woman 35 years old.

2. Obtaining gels containing labeled testosterone.

The thickener (polymer) was subjected to swelling overnight in the required amount of water. Labeled and unlabeled testosterone was dissolved in ethanol, if possible together with a permeation enhancer. The aqueous and alcoholic phases were mixed, then neutralized with triethanolamine. Mixing was carried out using a spatula until complete homogeneity. For percutaneous delivery, 17 formulations were evaluated. Compositions are given in table. 1 (in mass% relative to the total weight of the composition). Compositions 13 and 14 correspond to non-gel solutions.

Table 1

No. Testosterone Thickener 95% ethanol Triethanolamine RO OA Isopropyl myristate eleven one 1% Cbb. 934 R 58 1.35 0 0 0 12 0.5 1% Cbb. 934 R 58 1.35 0 0 0 thirteen 0.5 0 58 0 0 0 0 14 0.75 0 58 0 0 0 0 fifteen 2,5 1% Cbb. 934 R 58 1.35 0 0 0 sixteen 2,5 1% Cbb. 934 R 58 1.35 5 0 0 17 2,5 2% K1ise1 58 1.35 0 0 0 eighteen 2,5 2% K1ise1 58 1.35 5 0 0 nineteen 2,5 1% Cbb. 934 R 58 1.35 0 0 5 twenty 2,5 0.5% Cbb. 980 58 1.35 0 0 0 21 one 0.5% Cbb. 980 58 0.6 0 0 0 22 one 0.5% Cbb. 980 58 0.6 2 0 0 23 one 1% Cbb. 980 58 1.35 2 0 0 24 one 1% Cbb. 934 R 58 1.35 2 0 0 25 one 1% CAG. 934 R 58 1.35 2 2 0 26 one 0.5% Cbb. 980 58 0.5 one 0 0 27 one 0.5% Cbb. 980 58 0.5 one 0 0.5

(Cbb. = Cboro1®).

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3. The protocol of the kinetics of skin permeability.

3.1. Cell for the study of skin permeability.

Used static flow cells, including two unequal compartments (Fig. 1).

The upper cylindrical (cross-section: 2.54 cm ² ) donor compartment 1 allows you to apply the composition (in the form of a solution or in the form of a topical application) to the skin 2. This upper compartment 1 is connected to the lower compartment 4 using a metal ring.

The lower collecting compartment 4 (average volume 10 ml) also has an opening of 2.54 cm ² . The lateral channel 5, equipped with a needle and a catheter system, allows sampling. The homogenization of the medium in this lower compartment is carried out using a magnetic stirrer 3 (Fig. 1).

3.2. Biological membrane.

The skin of animals.

Biopsy samples of the skin of a male hairless rat (UR8 rat, 5 weeks) were obtained from the ventral surface of the abdomen. Adipose tissue localized from the inside was removed and the skin was cut to obtain samples with an area of more than 2.54 cm ² . Each sample was mounted on the lower compartment, directing the outer surface of the skin up.

Human skin.

The experiments were carried out on samples obtained after abdominal plastic surgery on the ventral surface of the abdomen in a 35-year-old white woman. Samples were used immediately after their preparation and cut into a dermatome with a thickness of about 250 μm.

3.3. Study Protocol

Two compartments 1 and 4 of the cell for studying skin permeability are connected using a fragment of skin 2. The capture compartment 4 is filled with a precisely measured volume of a 0.5% albumin solution. The cell is then placed for 15 hours in a thermostatic bath at 37 ° C, located on a mixing platform, until equilibrium between the skin and the capture medium occurs.

The kinetic measurement begins after applying the composition to skin 2. Apply about 20 mg to 2.54 cm ^{2 of} skin. After 24 hours, 2 ml of the sample was taken from the recovery compartment 4 at regular intervals and 2 ml of fresh albumin solution was immediately added to keep the volume constant. For each composition, 5 cells were used to study permeability. After storage in the cold, the samples were titrated by liquid scintillation.

4. Titration of C-testosterone.

The samples are divided into two fractions of 1 ml each, to which 4 ml of scintillation fluid (P1co G1iog 40®) is added, the measurement time is 5 minutes.

Samples are then titrated using liquid scintillation using a β spectrophotometer (Wesstap). In the scintillation process, quenching leads to a reduced detection sensitivity, which is accurately estimated by the established calibration curve, called the quenching curve. For each sample, the quenching-dependent measurement is calculated using an external standard and the true radioactivity is determined.

results

1. The solubility of testosterone.

A known amount of solvent (2 ml) is stirred at 37 ° C in a closed Erlenmeyer flask. Gradually add fractions, starting with a known amount of testosterone. The remaining amount of testosterone is weighed to determine the amount of testosterone in the Erlenmeyer flask (by difference).

Using the incremental method, the solubility of testosterone is determined at 37 ° C in water-ethanol mixtures and in the presence of various permeability enhancers, such as propylene glycol (PO), oleic acid (OA), propylene glycol dipelargonate (ROER), and Tgapci1o1.

table 2

Testosterone solubility in various carriers

Solvent PO (100%) RO / OA (50/50) OA (100%) ΡΟϋΡ (100%) Tgapsi1o1 (one hundred%) Solubility (mg / ml) 86 79 66 eighteen 140

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Testosterone solubility is also evaluated as a function of ethanol content.

Table 3

95% ethanol (w / w) Testosterone solubility mg / ml g / g 42/58 14 0.015 53/47 twenty 0,023 56/44 24 0,027 63/37 thirty 0,035 74/26 52 0,061 84/16 120 0.146 86/14 160 0.195 100/0 340 0.420

See graphs in FIG. 2.

2. The viscosity of the gels.

The rheological properties of the gels were characterized using a Sat-Meb controlled voltage rheometer, measuring parameters: plane / plane, diameter 2 cm, gap 1 mm, temperature 20 ° C.

The rheograms were depicted as a stream followed by a defined shear cycle:

ascending phase: voltage increase from 0 to 300 N / m ² for 2 min;

plateau: constant voltage 300 N / m ² for 1 min;

descending phase: undervoltage from 300 N / m ² to 0 within 2 min.

All gels were characterized by shear weakening (pseudoplasticity), without any hysteresis cycle.

Two rheological studies were carried out: the effect of the ethanol content on the viscosity of the Cagoro1®934 gel; the effect of the polymer content on the viscosity of Cagoro1®980 gel. 2.1. Viscosity of a water-alcohol gel Sagoro1®934P.

The effect of the ethanol content on the hydroalcoholic gels of Caglio1® is investigated by changing the content of 95% ethanol from 50 to 70%.

Table 4

Ethanol (wt.%) Viscosity (Pa) Maximum speed gradient (sec ' ¹ ) fifty 15.00 22.5 55 6,500 fifty 60 2,375 140 65 0,800 400

When the ethanol content exceeds 55%, the viscosity becomes less, with strong variations depending on the ethanol content. Viscosity increases with 50% ethanol.

2.2. The viscosity of the gel Sagoro® 980.

The effect of the polymer content on hydroalcoholic gels at 40% ethanol was studied by varying the content of Caboro® 980 from 0.3 to 0.7%.

Viscosity was measured on a plateau at a voltage of 300 N / m ² for 1 min.

Table 5

Sagoro1®980 (%) Viscosity (Pa) Maximum speed gradient (sec ' ¹ ) 0.3 0.125 1400 0.4 0.610 500 0.5 3,50 90 0.6 6.10 fifty 0.7 12 thirty

3. Skin permeability of testosterone compositions on rat skin.

The skin permeability of the compositions shown in table. 1, graft on rat skin was evaluated. Composition 25 cannot be evaluated due to heterogeneity (phase separation). The results of the amount of testosterone released after 24 hours and the rate of release after 24 hours are presented in table. 6.

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Table 6

Song Number Testosterone (%) The released amount (24 hours) (μg / cm ² ) Release Rate (24 hours) (%) eleven one 28.1 35 12 0.5 11.6 29th thirteen 0.5 24.4 61 14 0.75 36.7 61 fifteen 2.5. 16,2 8 sixteen 2,5 30.9 fifteen 17 2,5 10.5 5 eighteen 2,5 36.9 eighteen nineteen 2,5 44.5 22 twenty 2,5 18.5 nine / ^ 4 LL 4 L 21 one 21.0 2 / 22 one 33.5 42 23 one 34,4 43 24 one 36.9 46 25 one - - 26 one 30,0 37.5 27. one 62.0 77

Compositions 11-27: 8 mg / cm ² deposited.

4. Permeability of testosterone on human skin.

Compositions 26 and 27 were evaluated on human skin, together with composition 15 as a control. The results are detailed below.

Composition 26.

The average permeability parameters for 5 cells

Time (hour) Amount of C (mcg) 2 0 4 1.9 6 3,5 nine 5,6 12 7.4 24 15.3

The number / area of C / 8 (μg / cm ² ) Standard deviation Ts / 8 (p-5) 0 0 0.75 0.4 1.4 0.6 2.2 0.6 2.9 0.8 6.01 1,6

The average kinetic plot is shown in FIG. 3. Average flow: 0.26 μg / cm ² -h.

Composition 27.

The average permeability parameters for 5 cells

Time (hour) Amount of C (mcg) 2 0 4 3 6 5,6 nine 8.6 12 11.7 24 22.1

The number / area of C / 8 (μg / cm ² ) Standard deviation C / 8 (n = 5) 0 0 1,2 0.9 22.2 1,1 3.4 1,2 4.6 1.4 8.7 2,5

The average kinetic plot is shown in FIG. 4. Average flow: 0.39 μg / cm ² -h.

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Composition 15 (comparison).

The average permeability parameters for 5 cells

Time (hour) Amount (mcg) Quantity / area () / 8 (mcg / cm ² ) Standard deviation () / 8 (n = 5) '2 0 0 0 l t 1,1 And to m _? “Gi L Ί 6 2.2 0.85 0.5 nine 6.6 1.3 0.6 12 4.3 1.7 0.7 24 9.1 3.6 1,2

The average kinetic plot is shown in FIG. 5.

Average flow: 0.15 μg / cm ² -h.

Example 2

Materials and methods.

1. Materials.

[1,2,6,7- ³ H] -testosterone (Ategzyat aogaShpez): = 295, solution 1 mCi / ml (37 MBq / mL) in ethanol; specific activity 99 Ci / mol (3.66 TBq / mol), i.e. 336 mCi / mg (1-118).

Absolute ethanol SAG1O EGBA Ιοί Υ4Ν051154Ν

Materials Controlled by Laurenta Spes Wesz

Sagoro1®980

Ιοί 92010.4420

Sagoro1®934

Ιοί / 3898

Testosterone

Ιοί 92039/4461

Isopropyl myristate

Triethanolamine (TEM)

Propylene glycol (RSg)

10191169/5781

Ιοί 9304020/5721

Compositions for application to the skin (weight per 100 g of the total composition)

Composition Testostero n Thickener Absolute ethanol Neutralizing agent Propylene- ,glycol Isopropyl myristate 1 (cont.) 2,5 Sag. 934: 1.0 57 TEM: 1.35 - - η eleven floor. l / s \ /αιυ.Ήον. ν, υ and about 'TGG Λ. l ag \ l ± 2 / Ζ-γ. υ, υυ l with about _? and - 3 one Sag. 980: 0.8 58 TEM: 0.80 0.5 - 4 one Sag. 980: 0.9 67 NaOH (0.1 M): 4.725 0.5 5 one Sag. 980: 0.5 67 TEM: 0.50 - 0.5

Compositions are prepared as follows.

Composition 1.

g of radioactive gel is prepared as follows:

100 μl (100 μCi) of radioactive testosterone solution ( ³ N) is dried and resuspended with

595 mg of a solution containing 2.5 testosterone in 57 g of absolute ethanol;

391.5 mg of the main gel obtained by swelling 1 g of Cagoro1®934 in 38.15 g of water; after the final swelling and before use, the adhesive is homogenized for several seconds using a disintegrator and 1! ha Tigah;

13.5 mg of TEM.

The radioactivity concentration of the composition: 100 μCi / g of gel, i.e. 0.8 μCi and 200 μg of testosterone to apply 8 mg (about 10 μl) per cell area of 1.77 cm ² .

Composition 2.

g of radioactive gel is obtained in accordance with the following protocol:

100 μl (100 mCi) of radioactive testosterone solution ( ³ N) is dried and resuspended with

595 mg of a solution containing 1 testosterone and 0.5 g ΡΘ in 58 g of absolute ethanol;

393 mg of the main gel obtained by swelling 0.6 g of Cagoro1®980 in 38.7 g of water; after the final swelling and before use, the adhesive is homogenized for several seconds from to

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mg TEM, diluted in half with water.

The radioactivity concentration of the composition: 100 μCi / g of gel, i.e. 0.8 μCi and 80 μg testosterone to apply 8 mg (about 10 μl) per well.

Composition 3.

g of radioactive gel is obtained in accordance with the following protocol:

100 μl (100 μCi) of radioactive testosterone solution ( ³ N) is dried and resuspended with

595 mg of a solution containing 1 testosterone and 0.5 g of MS in 58 g of absolute ethanol;

398 mg of the main gel obtained by swelling 0.8 g of Cagoro1®980 in 38.1 g of water; after the final swelling and before use, the adhesive is homogenized for several seconds using the IIga Tigakh disintegrator;

mg TEM, diluted in half with water.

The radioactivity concentration of the composition: 100 μCi / g of gel, i.e. 0.8 μCi and 80 μg testosterone to apply 8 mg (about 10 μl) per well.

Composition 4.

g of radioactive gel is obtained in accordance with the following protocol:

100 μl (100 μCi) of radioactive testosterone solution ( ³ N) is dried and resuspended with

685 mg of a solution containing 1 testosterone and 0.5 g of isopropyl myristate in 67 g of absolute ethanol;

267.75 mg of the main gel obtained by swelling 0.9 g of Cagoro1®980 in 25.875 g of water; after the final swelling and before use, the adhesive is homogenized for several seconds using the IIg Tigah disintegrator;

47.25 mg No. 1OH (0.1 M).

The radioactivity concentration of the composition: 100 μCi / g of gel, i.e. 0.8 μCi and 80 μg testosterone to apply 8 mg (about 10 μl) per well.

Composition 5.

g of radioactive gel is obtained in accordance with the following protocol:

100 μl (100 μCi) of radioactive testosterone solution ( ³ N) is dried and resuspended with

685 mg of a solution containing 1 testosterone and 0.5 g of isopropyl myristate in 67 g of absolute ethanol;

305 mg of the main gel obtained by swelling 0.5 g of Cagoro1®980 in 30 g of water; after the final swelling and before use, the adhesive is homogenized for several seconds using the IIga Tigakh disintegrator;

mg TEM, diluted in half with water.

The radioactivity concentration of the composition: 100 μCi / g of gel, i.e. 0.8 μCi and 80 μg testosterone to apply 8 mg (about 10 μl) per well.

2. Methods.

Skin absorption ίη νίίΐΌ.

Principle.

Transdermal absorption η νίίΐΌ is studied quantitatively on human biopsy specimens obtained from the ventral surface of the abdomen and cut using a dermatome placed in a static diffusion cell (Franz cell), which makes it possible for the dermis to come into contact with the tissue experience medium (receptor medium), in which metered absorption through the skin.

Cell.

A skin biopsy is placed horizontally between the two parts of the cell, thus delimiting two compartments:

1) the epidermal compartment includes a glass cylinder having a strictly defined surface of 1.77 cm ² placed on top of the skin;

2) the skin compartment adjacent to the lower side of the cover includes a reservoir with a fixed volume having a side channel for sampling.

Two elements are interconnected by a clamp.

The lower compartment (dermal) is filled with tissue media made up of a solution of sodium chloride at a concentration of 9 g / l, supplemented with bovine serum albumin at a concentration of 15 g / l.

At each time point, the medium for experiencing tissue is completely taken from the lateral canal and replaced with a fresh medium.

The lower part of the cell is thermostated at 37 ° C. The homogeneity of temperature and content in the receptor medium is maintained by stirring (magnetic stirrer).

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The upper part (epidermal compartment) is open towards the outside, thus exposing the epidermal surface of the air in the laboratory.

Obtaining biopsy samples of the skin from the ventral surface of the human abdomen, cut using a dermatome.

These are samples of human skin from the ventral surface of the abdomen obtained as a result of plastic surgery from white-skinned donors. The skin is stored at -20 ° C before use. The adjacent subcutaneous fat is removed with a scalpel and the skin is cut with a thickness of about 0.5 mm using a dermatome. For each analysis, different skin samples are evenly distributed between batches.

The main protocol.

Franz cells are usually installed the day before application of the test composition. The epidermal compartment is in contact with the laboratory atmosphere, the skin compartment is thermostated at 37 ° C, and the skin is in contact with physiological serum containing albumin for about 17 hours. Under these conditions, the skin is well hydrated.

Using a micropipette, 10 μl of the solution (about 8 μl, about 8 μCi) is applied to the entire surface of the epidermis bounded by a glass cylinder. At certain points in time, after 2, 4, 6, 8 and 24 hours, samples of the fluid contained in the skin compartment are taken through the lateral channel. At each point in time, the tissue media is completely removed and replaced with fresh fluid.

At the end of the analysis, the treated skin surface is washed with 200 μl of various solvents in accordance with the following protocol:

1st washing: Ce1au1op® / Water (1/9, w / w);

2nd washing: water;

3rd washing: Ce1au1op® / water (1/9, w / w);

4th washing: water;

5th washing: water;

The application surface is then wiped with Ο-Ιίρ®.

Then, the epidermis and dermis are mechanically separated using a scalpel and dissolved in 1 and 3 ml of 8O1iepe® (Raskagb), respectively.

Measurement of radioactivity.

Detection was carried out using liquid scintillation using a particle counter Raskag6-1psagL 4530.

Receiving radioactive samples.

Tissue surviving medium, taken from the lower compartment of the diffusion cells, was placed directly in 15 ml of scintillation fluid (Ruoyogi 40K, Raskagb) and the radioactivity was measured.

Wash liquids and Ο-Ιίρδ® were placed in a vial containing about 30 ml of 95-degree ethanol and accurately weighed. After overnight incubation at 4 ° C, the radioactivity of precisely weighed aliquots from this dilution was determined and then the same procedure was carried out with tissue media.

The epidermis and dermis must first be dissolved in strong organic bases. They are dissolved for 24 hours at 37 ° C in 8O1EP 350® Raskagb (1 ml per 100 mg of tissue), to which 15 ml of scintillation fluid (Nyushs AIog 30®, Raskagb) is added and measurement is carried out.

Measurement of radioactivity.

Using the external calibration method, the normalization of the measurements is corrected, and quenching is also paid attention to in order to obtain the decay per minute (brt) count for the true activity of each sample. For each sample, subtract the background in cp. A specific quenching curve is established for each scintillation fluid.

The results are expressed in mass or percent of the substance found in the samples, relative to the amount introduced, which is determined by normalizing diluted suitably calibrators.

Transdermal absorption.

The transdermal absorption intensity was estimated by calculating the absorption in percent (%) of the applied amount (Οί) as a function of time% = (Ο1 / ςί) χ100, where 01 is the amount absorbed at time 1.

The average flow and the absorbed amount at each time interval were also determined and expressed respectively in ng / cm ² / I and in ng.

The averaged results correspond to 6-8 experimental determinations and are accompanied by the standard deviation values (86).

Comparison of the averaged results was performed using analysis of variance.

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3. Results.

Concentrations, expressed as percentages, represent the weight of the compound per 100 g of the total weight of the composition.

Permeability of testosterone ΐπ υϊϊγο in Franz cells (accumulated values).

Average values +/- 86 as a function of time

Composition Values Cell media environment 2 h 4h 6 h 8h 24 h one n = 6 % time 0.23% 0.42% 0.54% 0.63% 0.97% +/- 86 0.08% 0.12% 0.13% 0.15% 0.24% quantity (ng) 501 902 1184 1378 2113 +/- 86 169 254 278 322 518 2 n = 8 % time 0.40% 0.77% 1.03% 1.23% @ 1.98% @ +/- 86 0.18% 0.29% 0.36% 0.41% 0.59% quantity (ng) 351 666 892 1067 1721 +/- 86 159 256 317 360 508 3 n-8 % time 0.35% 0.72% 0.97% 1.17% 1.94% @ +/- 86 0.21% 0.34% 0.41% 0.47% 0.59% quantity (ng) 313 639 868 1047 1735 +/- 86 188 302 370 418 530 4 n = 7 % time 0.52% 0.94% @ 1.21% @ 1.40% @ 2.17% * +/- 86 ' 0.15% 0.27% 0.34% 0.38% 0.58% quantity (ng) 462 834 1071 1243 1927 +/- 86 133 237 301 341 514 5 n-7 % time 0.56% 1.04% @ 1.33% @ 1.53% @ 2.47% * +/- 86 0.49% 0.71% 0.78% 0.81% 0.99% quantity (ng) 484 899 1153 1324 2138 +/- 86 424 615 673 707 861

@: difference compared to control (analysis of variance, Fisher test, statistical significance p <5%).

*: difference compared to control (analysis of variance ΑΝΟνΑ, Fisher and Scheffe test, statistical significance p <5%).

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Corrected quantities (accumulated values).

Composition Values Quantities in 24 hours E ϋ E + 1) C + ϋ T one n = 6 % 36.35% 0.98% 37.33% 1.95% 47.03% 85.33% +/- 8ά 8.15% 0.55% 9.69% quantity (ig) 79072 2131 81203 4244 102306 185622 +/- 8ά 18516 1193 21075 2 n = 8 % 44.35% @ 1.55% 45.91% 3.53% 36.06% 83.95% +/- 84 8.44% 1.42% 5.23% quantity (ig) 38513 1349 39862 3069 31301 72844 +/- 84 7325 1235 4539 3 n = 8 % 40.56% 1.91% 42.47% 3.85% 41.47% 85.88% +/- 84 4.98% 1.02% 6.93% quantity (ng) 36200 1701 37900 3436 37015 76650 +/- 84 4443 916 6189 4 n = 7 % 44.56% 2.10% 46.66% 4.27% 31.77% 80.60% • +/- 84 5.37% 0.65% 4.99% quantity (ng) 39506 1865 41371 3792 28164 71462 +/- 84 4763 577 4419 5 n = 7 % 29.27% ° 2.97% * 32.24% 5.44% 46.58% 81.29% +/- 84 7.35% 1.18% 8.18% quantity (ng) 25393 2580 27973 4718 40397 70508 +/- 84 6374 1022 7096

E: epidermis; Ώ: dermis; C: cell; flushing; T: amount.

@: difference compared to control (analysis of variance, Fisher test, statistical significance p <5%).

°: difference with composition 2, 3 and 4 (analysis of variance, Fisher test, p <5%).

*: difference compared to control (analysis of variance ΑΝΟνΑ, Fisher and Scheffe test, statistical significance p <5%).

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Permeability of testosterone ίπ νίΐΐΌ in Franz cells (unaccumulated values). Average values +/- 8J as a function of time

Composition Values Cell media environment 2h 4 h 6 h 8 h 24 h one n = 6 % 0.23% 0.18% 0.13% 0.09% 0.34% +/- 8 (1 0.08% 0.05% 0.04% 0.03% 0.12% quantity (ng) 501 401 281 194 735 +/- 8ά 169 101 91 59 256 flow (ng / cm ² / b) 141.60 111.34 79.41 54.88 29.95 +/- 81 47.71 28.65 25.78 16.53 9.03 2 n = 8 % 0.40% 0.36% @ 0.26% @ 0.20% * 0.75% * +/- 8 (1 0.18% 0.13% 0.09% 0.07% 0.22% quantity (ng) 351 315 226 174 654 +/- 8ά 159 113 82 63 194 flow (ng / cm ² / b) 99.14 88.97 63.98 49.24 23.10 +/- 8ά 45.03 31.91 23.13 17.74 6.86 3 n = 8 % 0.35% 0.36% @ 0.26% @ 0.20% * 0.77% * +/- 8 (1 0.21% 0.15% 0.09% 0.07% 0.21% quantity (ng) 313 325 229 179 688 +/- 8ά 188 136 84 61 184 flow (ng / cm ² / b) 88.45 91.94 64.82 50.68 24.29 +/- 8 (1 53.09 38.33 23.86 17.25 6.49 4 n = 7 % 0.52% 0.42% @ 0.27% @ 0.19% @ 0.77% * . + / - 81 0.15% 0.15% 0.09% 0.06% 0.21% quantity (ng) 462 372 237 172 684 +/- 81 133 132 81 fifty 185 flow (ng / cm ² / b) 130.51 105.15 67.00 48.49 24.16 +/- 81 37.61 37.22 22.82 14.26 6.53 5 n = 7 % 0.5b% @ 0.48% * 0.29% * 0.20% (o} 0.94% * +/- 81 0.49% 0.23% 0.08% 0.05% 0.20% quantity (ng) 484 415 254 171 814 +/- 81 424 198 73 46 177 flow (ng / cm / b) 136.86 117.14 71.74 48.22 28.53 +/- 81 116.80 55.98 20.67 13.04 6.26

@: difference compared to control (analysis of variance, Fisher test, statistical significance p <5%).

*: difference compared to control (analysis of variance ΑΝΟνΑ, Fisher and Scheffe test, statistical significance p <5%).

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Claims (14)

CLAIM 1. A pharmaceutical composition comprising from 0.5 to 5.0% (w / w) testosterone; from 40.0 to 75.0% (w / w) of at least one C ₂ -C ₆ alcohol; from 0.1 to 5.0% (w / w) of at least one thickener; from 0.1 to 5.0% (w / w) propylene glycol; from 0.05 to 5.0% (w / w) isopropyl myristate. 2. A pharmaceutical composition further comprising water. 3. The pharmaceutical composition according to claim 1, in which said C ₂ -C ₆ alcohol is selected from the group consisting of ethanol, propan-1-ol and propan-2-ol, and mixtures thereof. 4. The pharmaceutical composition according to any one of claims 1 to 3, in which said thickener is selected from the group consisting of polymers based on acrylic acid, carboxymethylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, cellulosic polymers, and mixtures thereof. 5. The pharmaceutical composition according to any one of claims 1 to 4, in which the specified thickener includes at least one thickener selected from the group consisting of carbomers, for example Sagor1® 980, Sagor1® 934P, Sagor1® 1342, Sagor1® 1382, and their mixtures. 6. The pharmaceutical composition according to any one of claims 1 to 5, further comprising a base, preferably selected from the group consisting of triethanolamine, sodium hydroxide, ammonium hydroxide, potassium hydroxide, arginine, aminomethyl propanol, tromethamine, and mixtures thereof. 7. A gel useful for transdermal delivery, comprising a pharmaceutical composition according to any one of claims 1 to 6. 8. Dosage packaging, which contains a pharmaceutical composition according to any one of claims 1 to 6 or the gel according to claim 7. 9. Dosed packaging of claim 8, which is a single metered packaging or packaging, designed for several receptions. 10. A pump dispenser containing the pharmaceutical composition according to any one of claims 1 to 6 or the gel according to claim 7. 11. The dispenser of claim 10, in which the pump is a manual pump. 12. A method of obtaining a pharmaceutical composition according to any one of claims 1 to 6 or gel according to claim 7, comprising the following stages: obtaining a mixture containing at least one C2-6 alcohol, propylene glycol, isopropyl myristate and testosterone; adding a thickener to the mixture and mixing. 13. The method of claim 12, further comprising the step of adding water to the mixture and re-stirring. 14. The method of claim 12, further comprising the step of adding a base to the mixture and re-mixing.