Testicular Preparations

TESTICULAR PREPARATIONS, organ preparations obtained from the sex glands (testicles) of animals, namely: dried and powdered or made into tablets and lozenges testicles, dry or liquid extracts from them (extraction with physiological solution, alcohol, glycerin, etc.), so-called testicular fluid according to N. P. Kravkov (Ringer-Locke solution that has passed through the vessels of isolated testicles), products of artificial digestion of testicular tissue (optones Abderhalden, lysates Tushnov, etc.) and similar preparations (see Organ preparations). Preparations of the spermine type are also included (see Spermine). Chemically, testicular preparations represent a complex of various substances, the content of which varies depending on the nature of the raw material and methods of its processing. Various authors have found in testicular preparations nucleoproteins, peptides, various amino acids (arginine, histidine, tyrosine, tryptophan, cystine, leucine, etc.), purine bases (adenine, guanine, hypoxanthine, xanthine), creatine, enzymes, phosphorus (up to 1.2-1.5% in dried testicles), a relatively high content of zinc (up to 32 mg per 100 g of dry substance) and others. Biologically, some of these substances, especially with parenteral administration, are active, but not specific to testicles, as they are found in organ preparations obtained from other organs. The male sex hormone (Androkinin, Andotin) has not been proven in the testicular preparations mentioned; it is contained in preparations obtained in recent years by special processing of testicles, blood or urine of men (see below). Testicular preparations can be divided into two large groups: those not containing and those containing the male sex hormone. The preparations of the first group (Orchidin, Orchicrin, etc.) for internal and subcutaneous use are mainly on the market. The beginning of the scientific study of preparations of the first group was laid by the experiments of Brown-Séquard (1889) and was the impetus for numerous experiments on animals and humans, which clarified the pharmacodynamic features of such preparations. Only individual authors reported positive results of replacement therapy (see Organ preparations) with the help of testicular preparations. Nussbaum (1908) obtained in castrated male frogs restoration of the grasping reflex (much more vividly this was obtained by Stemacn, when introducing extract from nervous tissue), thickening of the thumb of the forelimbs (the same is given by ovarian substance) and hypertrophy of their muscles. Levi described better growth of the comb and wattles in capons fed with testicular substance than in control birds. Later Smith and Crocker, under the influence of subcutaneous injections of saline extracts from testicles, obtained in chickens increase and greater brilliance of the comb and wattles, greater brilliance of feathers, decrease in egg-laying, and some chickens tried to cover others like roosters. Other authors, however, did not confirm this. When applied to humans, the results of replacement therapy were contradictory. Most authors (Walker, Borchardt, Berkeley, etc.) came to the conclusion that the sometimes occurring increase in libido sexualis is due to the action of the type of nonspecific parenteral therapy, and not the hormonal effect. Therefore, no effect is observed in eunuchs. Some clinicians explain this by the fact that for full compensation of the phenomena of falling out after castration, the introduction of one sex hormone is still insufficient, since sexual development and sexual characteristics also depend on other endocrine glands (anterior pituitary, adrenal cortex, etc.). More consistent data were obtained regarding the tonic ('dynamogenic,' according to Brown-Séquard) action of testicular preparations—increase in the ability to muscular work, measured by an ergograph, more rapid recovery from muscular and nervous fatigue, increased excitability of the spinal cord centers, etc. However, experimentally (Vincent), similar results could also be obtained with extracts from other organs. The metabolism lowered after castration under the influence of testicular preparations clearly increases; with a normal level of metabolism, the effect is less noticeable. The same result was obtained with the introduction of preparations from ovaries. In relation to the cardiovascular system, a tonic action was also observed, weakly expressed in humans. Experimentally, the amplitude increased and the rhythm of the isolated heart of warm-blooded animals became more frequent; with intravenous administration, blood pressure significantly decreased, expansion of the coronary vessels was noted, and so on. Spermine and other organ preparations have the same effect. With repeated injections of large doses of testicular preparations, atrophy of the testicles and ovaries was observed, sometimes with complete sterilization of females and males. This kind of action is apparently not hormonal, but immunocytotoxic in nature (Bayer). In recent years, it has been possible to obtain preparations containing the male sex hormone (in 1929 Funk and Harrow and independently of them Gallagher, Koch, McGee and Moore). The purification of the hormone was so successful that it was also obtained in crystalline form (Frattini and Maino, 1930; Butenandt, 1931). Chemically, it is an oxiketone, having the composition C₁₉H₂₈O₂. Its hydroxyl has the character of a secondary alcohol, not a phenol; therefore it must be considered as a saturated system (Scheller). Chemically, it is similar to the female sex hormone (extracted with chloroform, forms alkaline salts, is contained in the fraction of fatty acid), but according to Laquer (Laquer with colleagues, 1931) it has not an acidic, but rather a basic character. Soluble in various organic solvents, very resistant to temperature, medium acids and alkalis, can be distilled with the removal of phenolic impurities without loss of activity. In suitable solutions, it is stable for more than a month (Loewe, Voss). It is contained in large quantities in the urine of men, in much smaller quantities in the blood, testicles and their appendages; it has also been found in the male catkins of willow (Salix caprea) and other plants. Most preparations are obtained from the urine of men (e.g. Hombreol and others). The effect is best manifested with subcutaneous or intramuscular administration; when taken orally, at least 10 times the dose is required (Koch). Some pain at the injection site of the preparation is noted. Preparations of this kind have specific (hormonal) activity, which is evident from their effect on secondary sexual characteristics and accessory gonads in castrated roosters, mice, rats and guinea pigs. A few days after castration of roosters, their comb and wattles undergo reverse development, become pale; then the comb falls to one side, looks wrinkled, desquamation of the epithelium occurs. A few days after injection of the hormonal preparation, redness of the comb, increased turgor and increase in size occur. After the injections are stopped, the reverse development of the comb size quickly occurs. This reaction is used for the quantitative expression of the biological activity of preparations. As a unit of action (1 rooster unit), Funk with colleagues (1930) take the smallest amount of preparation that causes an increase in the size of the comb and wattles by 10 mm within 10 days. Laquer with colleagues (1932) take 1/8 of the smallest dose (divided into 8 subcutaneous or intramuscular injections over 4 days) as 1 rooster unit, which on the fifth day after the first injection in most roosters causes a minimum growth of the comb by 15%. As long as we have no other substances that cause such an effect in castrated roosters. Testicular hormonal preparations have a bright effect on the seminal vesicles of rodents. After castration, these glands decrease in size and weight, contain almost no secretion, are histologically sharply changed. After injection of a preparation containing the male sex hormone, the size and weight of the seminal vesicles significantly increase, the secretion in their lumen becomes abundant and well-staining, instead of flattened cubic epithelium, a high-cylindrical, cell-rich epithelium is found, secretory granules that were absent appear, the Golgi apparatus, which was strongly wrinkled, develops well, instead of chondriosomes that had disintegrated into lumps, they acquire the appearance of long threads. The phenomena described by Loewe and Voss (1929) they called 'cytologische Regenerationstest'. Their data have been confirmed by other authors (Martin, Rocha and Silva for mice, Moore and Gallagher for rats, etc.). As one mouse unit of activity of testicular preparations, Loewe and Voss take the smallest amount of preparation which, divided into 8 injections, on the 10th day gives the described cytological changes. The ratio between different biological units according to various authors is approximately as follows: 1 rooster unit = 10 mouse or 5 rat units. For complete restoration of the secondary sexual characteristics of a capon, it is necessary to introduce up to 60 rooster units per day, obtained from approximately 6 kg of bull testicles (Laquer, 1932). In accordance with this, it is believed that there is no reserve of hormone in the testicles; it is possible that modern methods of its extraction are still very imperfect.

Hormonal preparations also prevent the enlargement of the prostate gland in rats after castration or even restore the gland to normal (Moore and colleagues). In humans with prostate hypertrophy, after several (15-40) daily injections of the preparation Hombreol (1 cc³ = 4 rooster units), a reduction in gland size and improvement in the patients' general condition was observed (Cappellen, 1933). Data from some authors (McCullagh, 1932, and others) show that the testicles contain at least two hormones: the hormone under consideration (Androtin, Androkinin), which does not prevent the hypertrophy of the pituitary and adrenal glands in rats that usually occurs after castration, and another hormone (Inhibin), which can be separately obtained from the testicles and whose deficiency in production in old age in humans is said to explain the hypertrophy of the prostate gland. It is possible that the action of the hormone is not direct, but through stimulation of the function of the anterior pituitary lobe, since experiments show that in hypophysectomized male rats, preparations with sex hormone do not cause changes in the weight and histological structure of the testicles, prostate gland, and seminal vesicles. In the treatment of eunuchs and eunuchs with hormonal T. p., the appearance of a positive reaction to the anterior pituitary hormone in the blood of patients was noted (McCullagh and colleagues, 1933). In these same patients, with prolonged treatment (2-3 months, injections every 2-3 days with 6 rooster units), an increase in libido sexualis, a decrease in vascular and nervous disorders, a general increase in energy, the appearance of nocturnal emissions, the appearance of motile spermatozoa in prostatic fluid, a decrease in cholesterol in the blood, a slight increase in basal metabolism, and the appearance of the hormone in the urine were noted (in 7 out of 19 cases), which was not the case before treatment. Research showed that a single injection of the hormone is excreted in the urine within three days, which is in agreement with the data from animals. Complete restoration of secondary sexual characteristics in humans with sharply reduced or absent function of the sex glands has not yet been achieved. The hormone also affects females: suppression of the sexual cycle in rats, cessation of estrus phenomena (Lendle, Ihrke and D'Amour, 1931, and others), growth of the clitoris in guinea pigs (de Jongh and Mulder, 1932), an increase in the size of the comb in normal or castrated chickens (Freud, de Jongh, Laqueur and Munch, 1930) has been established. Clinical observations on hormonal T. p. (mostly of laboratory preparation) are still insufficient to judge the necessary dosage for humans, duration of administration, indications, and results of therapy. The general indication for the use of T. p. is all cases of insufficiency of testicular secretion (e.g., after their destruction by trauma, tuberculosis, gummas), disorder of libido sexualis and potentia coeundi. In accordance with the views on the physiological role of the testicles, T. p. were tried to be applied in senile marasmus, arteriosclerosis, functional neuroses, cardiac weakness, metabolic diseases (gout, obesity), in some intoxications and auto-intoxications, etc. The dosage of preparations not containing the hormone is purely empirical—liquid preparations 20-30 drops per dose several times a day or 1-2 cc³ under the skin daily or every other day, dry preparations 0.5-1.0 per dose several times a day. Before the discovery of the male sex hormone, a positive therapeutic effect from large doses of dry preparations was hoped for, however, cases where psychotherapy was excluded did not confirm this.