HK1068003B - Pharmaceutical composition comprising gamma-butyrobetaine - Google Patents

Description

Pharmaceutical composition containing gamma-trimethylammoniumbucillus salt

no marking

The invention relates to a known medicament and a pharmaceutical composition containing the same for further medical use, in particular for normalizing and stimulating sexual activity and sexual potency in mammals. The present invention discloses the novel utility of several known substances which exhibit unexpected pharmacological activity when combined. In particular, a pharmaceutical composition is disclosed which contains as active ingredients gamma-trimethylammoniumbutol salt (GBB) and 3- (2, 2, 2-trimethylhydrazinium) propionate (THP) or a phosphodiesterase inhibitor.

GBB (actinine), an intermediate in the synthesis of mammalian carnitine, was originally described as a toxic substance inducing tachypnea, sialorrhea and lacrimation, mydriasis, vasoconstriction and cardiac arrest (Linneweh W.Z physiol chem., 1929; 42: 181). Further studies have shown very low toxicity of GBB (LD50 7000mg/kgSubc.) (Rotzsch W, Lorenz I, Strack E.acta biol med ger 1959; 3: 28-36). The cardiovascular effects of GBB were compared to acetylcholine (HoseineA, McLennan H. pharmaceutical action of gamma-butyrobetaine. Nature 1959; 183: 328), but this data was later overturned by the same authors who in fact studied the effects of GBB methyl ester. GBB is considered pharmacologically inert by other investigators (Hosein EA, Proulx P.isolation and basic functions of beta estisers in braine metabolism.Nature 1960; 187: 321. BurrgenASV, Hobiger F.Brit J Pharmacol.1949; 4: 229.Strack E, Foesterling K.Z physiol chem.1, 953; 295: 377). In contrast, GBB was later shown to have free radical scavenger function (Akahira M, Hara A, Abiko Y. Effect of MET-88, a gamma-butyl beta hydrolase inhibitor, on myocardial derivatives induced by a hydrogen peroxide in the isolated treated heart. Fundam Clin Pharmacol.1997; 11 (4): 356) and to protect cardiac activity (Kalvins I, Veveris M. Latvian patent N.11727). Pharmaceutical compositions containing GBB as an active ingredient have also been found to be useful in the treatment of carnitine deficiency (Cavazza c. pharmaceutical composition comprising gamma-butyrobetaine. uk Patent Application GB 2091101 (1982)). No data are reported on the effect of GBB on sexual activity and performance in mammals.

3- (2, 2, 2-trimethylhydrazinium) propionate (THP) is also known as a Mildronate or Quaterine (British patent 2105992) drug. It can interfere with carnitine biosynthesis and thus limit the transport of long-chain fatty acids through the mitochondrial membrane (SimkhovichBZ, ShutenkoZV, Meirena DV et al.3- (2, 2, 2-trimethyo) propionate (THP) -a novel-butylbetaine inhibitor with chiral protective properties. biochem Pharmacol 1988; 37: 195). It has therefore found its application as a metabolic corrector in ischemic diseases of different origins and as a cytoprotective agent under hypoxic conditions.

The Lasevia patent LV11728 discloses a pharmaceutical composition comprising 3- (2, 2, 2-trimethylhydrazinium) propionate and gamma-trimethylammoniumbucillus salts for the treatment of cardiovascular diseases.

However, there is no data on the effect of 3- (2, 2, 2-trimethylhydrazinium) propionate (THP) or its use in combination with other substances on sexual activity and performance in mammals.

We have surprisingly found that gamma-trimethylammoniumbucillus and/or THP induces a substantial and long lasting increase in sexual activity in experimental animals. Moreover, the combination of these two substances produces a more durable and higher level of intracavernosal pressure increase than either component alone. Furthermore, GBB or its use in combination with THP has a positive effect on the cavernous pressure induced by the reflex stimulation. Thus we have surprisingly found that GBB, or its use in combination with THP, helps to stimulate sexual activity and performance in mammals. This activity cannot be attributed to the known effects of GBB and/or THP on fatty acid turnover or other known physiological effects of said substances.

Pharmacological effects on mammal sexual activity of GBB, THP and combinations thereof were performed by model studies of sexual intercourse behavior in rats under physiological depression.

The experiment was performed with adult Wistar rats of both initial sexes, starting at a body weight of 300-. During the experiment, six animals per group were housed in standard cages. The feed was a standardized food R70 (labcor, LactaminAB, sweden). Room temperature was maintained at 21-23 ℃, relative humidity was maintained at 65+ 10%, 12 hours light/dark cycle. The average water consumption of the rats was determined to be 2-12% (average-10%) of their body weight one week prior to the experiment.

Male rats were randomly divided into 4 groups of 6 rats each and supplied with the following aqueous solutions for 6 weeks:

group 1 (control group) - - -drinking water without additives;

group 2 (GBB group) -drinking water with added gamma-trimethylammoniumbutol (0.015% by weight) resulted in an average daily intake of 15 mg/kg gamma-trimethylammoniumbutol;

group 3 (THP group) -drinking water with THP (0.06% by weight) added resulted in an average daily intake of 60 mg/kg THP;

group 4 (GBB + THP group) -water with addition of THP (0.06% by weight) and gamma-trimethylammoniumbutol (0.015% by weight) resulted in an average daily intake of 60 mg/kg THP and 15 mg/kg gamma-trimethylammoniumbutol.

Sexual intercourse activity was tested 4 times in male rats: after 1 week, 4 weeks, 6 weeks and 48-50 hours after cessation of drug intake, all animals received drinking water without additives.

The test was performed at 10-12 am. Each group of 6 male rats was placed in a clean, well-lit cage. After an adaptation period of 5 minutes, 2 female rats were placed in the cage for 10 minutes. The following data were collected for each male rat:

1) sexual intercourse intensity (number of intercourse times during exposure period);

2) wake-up periods, recording the delay time respectively-the period during which the male rats showed interest in the female rats; and the number of approach/ride attempts during the exposure period.

3) Post-coital period-behavior of male rats within 5 minutes after removal of female rats. Post-coital behavior is described by the following markers: 0- -animal negative, lying down; 1- -animals are quiet and hair is cleaned; 2- -animal unstable, estrus; 3- -animal is active and good at fighting.

The female rats used were in estrus and were induced by an intraperitoneal injection of 0.2ml of 0.1% estradiol dipropionate 48 hours prior to the test.

Although the sexual behavior of the rats in the experimental group was significantly different from that of the control group, their water consumption did not show a substantial change due to the experimental material.

One week after the start of the experiment, the rats receiving GBB or GBB + THP showed substantially higher sexual interest and activity in sexual contact, as well as longer post-coital agonism. Continued use of GBB resulted in increased sexual activity, with the response being in terms of higher sexual intensity, while the oestrus versus overall activity of the animals was less affected (tables 1-4).

TABLE 1 Effect of pharmaceutical compositions on the number of riding attempts in male rats

Duration of administration	1 week	4 weeks	6 weeks	After administration
					Control	1.8±0.8	2.2±0.5	2.3±0.5	2.7±0.5
GBB	3.8*±0.4	3.7±0.7	3.3±0.6	3.2±0.5
					THP	2.7±0.6	3.0±0.7	3.7±0.5	3.4±0.7
THP+GBB	3.8*±0.4	4.0*±0.4	3.7±0.5	4.2*±0.4

P < 0.05) compared to control

TABLE 2 Effect of pharmaceutical compositions on Pre-ride delay time (minutes)

Duration of administration	1 week	4 weeks	6 weeks	After administration
					Control	5.8±1.4	3.8±0.8	4.7±1.1	3.5±0.7
GBB	3.7±0.9	1.8*±0.4	2.8±0.9	2.6±0.6
					THP	5.3±1.0	2.1±0.5	2.3±0.5	2.4±0.7
THP+GBB	3.5±0.6	1.6*±0.4	1.8*±0.4	1.7*±0.4

P < 0.05) compared to control

TABLE 3 Effect of pharmaceutical compositions on the number of intercourse

Duration of administration	1 week	4Week (week)	6 weeks	After administration
					Control	0.3±0.3	0.5±0.3	0.5±0.2	0.5±0.2
GBB	0.8±0.3	1.3±0.5	1.2*±0.2	0.7±0.2
					THP	0.5±0.3	1.2±0.4	0.8±0.3	1.0±0.3
THP+GBB	0.8±0.3	1.8*±0.4	1.5±0.4	1.2*±0.2

P < 0.05) compared to control

TABLE 4 Effect of pharmaceutical compositions on post-coital agonism in rats

Duration of administration	1 week	4 weeks	6 weeks	After administration
					Control	0.8±0.3	1.0±0.4	1.2±0.3	1.5±0.4
GBB	2.0*±0.4	1.2±0.3	1.5±0.4	1.2±0.3
					THP	1.0±0.4	1.4±0.5	1.8±0.3	1.2±0.4
THP+GBB	2.0*±0.4	1.4±0.5	1.7±0.3	1.8±0.4

P < 0.05) compared to control

The combined use of GBB and THP elicits an increase in sexual interest and sexual activity throughout the experiment. After discontinuation of dosing, only the GBB + THP group showed higher intercourse activity compared to the control group.

We thus experimentally demonstrated that GBB alone and GBB in combination with THP, after a treatment period of 6 weeks, produced considerable and durable sexual activity in male rats. Furthermore, the method is simple. We have found that: the combined use of the two substances has a surprising increase in potency compared to their use alone.

In further experiments, the novel compositions were compared to a known sexual performance stimulant papaverine (Sarosdy MF, Hudnall CH, Erickson DR, Hardin TC, Novicki DE.A. productive double-blind triple of intracorporeal papaverine Verstagladin E1 in great effort of erectile. J Urol, 1989; 141: 551), which is an effective erection stimulant in cavernous body injections.

The experiment was performed with adult male rats weighing 300-410 g. The effect of the test material on penile erection was evaluated using an experimental model measuring changes in cavernous pressure (Chen KK et al. J Urol, 1992; 147: 1124).

Rats were anesthetized with sodium pentobarbital (intraperitoneal injection 50 mg/kg + intravenous injection 8 mg/kg/h). Body temperature was maintained at 37-37.4 deg.C (rectal control) with a heating lamp. The airway is inserted into the trachea to ensure adequate breathing under anesthesia. A25 gauge needle filled with heparinized saline was connected to a pressure transducer and then inserted into the corpora cavernosa. The cavernous pressure and type II standard leads in the ECG were recorded continuously with physiograph DMP-48(Narco Bio-Systems, USA). Some experiments also recorded arterial pressure of the common carotid artery. The effect of the test material was determined by both intravenous and cavernous routes of administration. For injection of the sponge, the material was dissolved in an isotonic (0.9%) sodium chloride solution and injected in a dose of 0.05ml liquid. Papaverine hydrochloride, clinically used for sexual potency testing, was used as a positive standard (0.2 mg per rat in cavernous injection, 2.0 mg per kg in intravenous injection). Gamma-trimethylammoniumbutol salt (GBB) alone and in combination with THP or a phosphodiesterase inhibitor, particularly sildenafil.

Gamma-trimethylammoniumbutol (GBB) (intracavernosal injection 0.02-0.1 mg/rat, usually 0.05 mg/rat; intravenous injection 2.0 mg/kg) and THP (intracavernosal injection 0.2 mg/rat; intravenous injection 10.0 mg/kg) were introduced separately, and in combination.

Sildenafil (cavernous injection 0.15 mg/rat; intravenous injection 3.0 mg/kg) alone and in combination (GBB + sildenafil).

It was found experimentally that cavernous injection of GBB produced a significant dose-dependent, but relatively short-acting, increase in cavernous pressure (table 5).

TABLE 5 influence of Spongilla injection drugs on Spongilla pressure in anesthetized rats

Medicine	Dosage form	Increase in cavernous body pressure		Duration of effect
						Milligrams of	Millimeter mercury column	Papaverine%	Minute (min)
GBB	0.02	11.25**±3.3	30.6	3.0*±0.7
					GBB	0.05	31.5±5.1	85.7	4.3*±0.9
THP	0.2	2.7**±1.5	7.3	0.8**±0.4
					THP+GBB	0.2+0.05	40.0±5.6	108.8	10.4±2.0
Sildenafil	0.15	38.5±7.3	104.8	7.5±2.7
					Sildenafil + GBB	0.15+0.05	35.2±8.4	95.8	17.6*±4.3
Papaverine	0.2	36.75±4.1	100	8.8±1.4

P < 0.05 x) compared to papaverine p < 0.01 compared to papaverine

Increase in papaverine production, expressed as%

THP does not cause significant changes in the internal pressure of the sponge. GBB is also less active than papaverine in this assay. Unexpectedly, the effect of GBB in combination with THP or sildenafil is comparable to or superior to that of papaverine. The effect and duration of the combination are superior to the effect induced by each component alone.

Since cavernous injections are not commonly used due to inconvenience to the patient, the intravenous route of administration is used for further evaluation.

Experiments have shown that intravenous administration of papaverine and THP has little effect on the cavernous pressure, while GBB and GBB-THP are very effective in increasing the cavernous pressure (Table 6).

It is noteworthy that GBB-THP combination and GBB + sildenafil combination maintained the effect 2.25 times longer than GBB alone, and even 5.46 times longer accordingly. Also noteworthy are: only GBB-THP combination induced a significant positive response to reflex penile stimulation, resulting in increased cavernous body pressure, which is atypical for anesthetized animals.

TABLE 6 Effect of intravenous injection of pharmaceutical compositions on cavernous body pressure in anesthetized rats

Pharmaceutical composition	GBB	THP	THP+GBB	Sildenafil	Sildenafil + GBB	Papaverine
							Reflectivity enhancement of sponge pressure (mmHg)	7.3*±2.0	2.7±1.2	21.7*±10.4	11.8*±3.6	7.3*±2.1	0.3±0.3
Variation of sponge pressure (mm Hg)	22.0*±3.6	1.3±0.9	29.7**±4.3	12.3*±4.8	28.4*±7.9	0.6±0.6
							Duration of Effect (minutes)	2.8±0.7	0.9±0.5	6.3**±1.9	2.5±1.1	15.3*±4.2	0.9±0.9

P < 0.05 compared to papaverine

P < 0.01) compared to papaverine

Thus, it was confirmed that the pharmaceutical composition containing GBB or its combination with THP or sildenafil not only resulted in an increase in the sponge pressure upon sponge injection, but also was effective upon intravenous administration, as opposed to papaverine. We have demonstrated that compositions containing GBB + THP and GBB + sildenafil, compared to each component used alone, have surprising effects and unexpected duration of effects in inducing an increase in cavernous pressure, and are able to restore a positive reflex response to mechanical penile stimulation.

Given the positive effects these substances exhibit when administered orally, whether introduced parenterally or not, they are helpful in stimulating sexual activity and in stimulating an erection in normal and physiologically depressed states.

In the case where the active ingredient is administered by parenteral injection or orally as drops, syrup or beverage, the pharmaceutical composition contains GBB + THP, or GBB + sildenafil in a total amount of 0.5 to 40% by weight of the total weight of the pharmaceutical form and distilled water, physiological saline, a glucose solution, or a buffer solution as a pharmaceutically acceptable solvent.

In the case of the combined administration of the active ingredients as a tablet, caplet, capsule, pill, granule or powder, the pharmaceutical composition contains GBB + THP or GBB + sildenafil in a total amount of 0.5 to 5 grams per tablet, caplet, capsule, pill, granule or powder dosage unit.

In the case of transdermal, topical, sublingual, urethral or intranasal administration of the active ingredient, the content thereof is 0.5-40% by weight of the total weight of the pharmaceutical form.

In addition, the pharmaceutical compositions may contain other pharmaceutical agents, for example other phosphodiesterase type V inhibitors (vardenafil, tadalafil and related substances).

Claims (4)

1. Use of a pharmaceutical composition comprising gamma-trimethylammoniumbucillus salt as a free base or a pharmaceutically acceptable salt thereof, or a combination of gamma-trimethylammoniumbucillus salt and a drug for stimulating sexual activity in a mammal for the manufacture of a medicament for normalizing and stimulating sexual activity and sexual potency in a mammal; wherein the drug for stimulating the sexual activity of the mammal is selected from the group consisting of: as a free base or a pharmaceutically acceptable salt, 3- (2, 2, 2-trimethylhydrazinium) -propionate, a type V phosphodiesterase inhibitor.

2. The use of claim 1, wherein the phosphodiesterase type V inhibitor is selected from the group consisting of: sildenafil, vardenafil, tadalafil.

3. The use of claim 2, wherein the phosphodiesterase type V inhibitor is sildenafil.

4. The use of any preceding claim, wherein the composition further comprises a pharmaceutically acceptable diluent or carrier.