阅读说明：本技术 吡咯喹啉醌或其盐在制备用于防治前列腺增生药物中的用途及药物组合物 (Application of pyrroloquinoline quinone or salt thereof in preparation of drugs for preventing and treating prostatic hyperplasia and pharmaceutical composition ) 是由 王广基 甄乐 王健鲲 于 2021-01-15 设计创作，主要内容包括：本发明公开了吡咯喹啉醌或其盐在制备用于防治前列腺增生药物中的用途及药物组合物。吡咯喹啉醌或其盐对前列腺增生具有显著的抑制作用。(The invention discloses application of pyrroloquinoline quinone or a salt thereof in preparing a medicament for preventing and treating prostatic hyperplasia and a medicinal composition. The pyrroloquinoline quinone or the salt thereof has a remarkable inhibition effect on the prostatic hyperplasia.)

1. Application of pyrroloquinoline quinone or salt thereof in preparing medicaments for preventing and treating prostatic hyperplasia.

2. Use according to claim 1, characterized in that: said pyrroloquinoline quinone or salt thereof is administered by enteral or parenteral or topical route.

3. Use according to claim 1, characterized in that: the administration route of the pyrroloquinoline quinone or the salt thereof is oral, sublingual, transdermal, intramuscular or subcutaneous, skin mucosa, vein and urethra.

4. A pharmaceutical composition for preventing and treating prostatic hyperplasia, which is characterized in that: comprises pyrroloquinoline quinone or a salt thereof in a prophylactically or therapeutically effective amount.

5. The pharmaceutical composition of claim 4, wherein: contains 0.1-99.9 wt% of pyrroloquinoline quinone or a salt thereof as an active ingredient.

Technical Field

The invention relates to the field of medicines, in particular to application of pyrroloquinoline quinone or a salt thereof in preparing a medicine for preventing and treating prostatic hyperplasia and a pharmaceutical composition.

Background

Benign Prostatic Hyperplasia (BPH) has become the most common progressive disease causing micturition disorders in middle-aged and elderly men, gradually showing clinical symptoms mostly after age 40, with a prevalence rate of 42-90% in men over age 45 and increasing year by year as age increases, up to 70-80% by age 80. BPH is mainly clinically manifested as prostate volume increase and lower urinary tract symptoms, and seriously affects the life quality and physical and psychological health of middle-aged and elderly people. The actions of androgens and their downstream related cytokines are thought to be the major cause of disruption of prostate homeostasis and resulting hyperproliferation of prostate tissue cells.

The current treatment modalities for BPH are mainly divided into surgical resection and drug therapy. The problem of the patient's aging causes great limitation to the surgical treatment and is easy to cause various complications; in addition, the prostate gland produces multiple immunoglobulins, and has a local immune function that protects the reproductive system from bacterial and other pathogenic microorganisms, which should be retained as much as possible. Drug therapy is often limited to α 1 receptor blockers, 5 α reductase inhibitors, or a combination of both; and the curative effect is gradually reduced along with the treatment and the duration of the disease. Therefore, it is of great interest to develop novel drug molecules for the prevention or treatment of BPH.

PQQ (pyrroloquinoline quinone) with molecular formula of C₁₄H₆N₂O₈Molecular weight is 330.21, and its structural formula is shown below. In recent years, pyrroloquinoline quinone has been shown to be an important nutrient for mammals, and has a wide range of physiological and pharmacological activities, including growth promoting, antioxidant, antidiabetic, and neuroprotective effects; more notably, pyrroloquinoline quinone also has good safety (Bioscience, Biotechnology, and Biochemistry, 2016, 80, 13-22). However, there is no report on pyrroloquinoline quinone or a salt thereof for the prevention or treatment of BPH.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to provide application of pyrroloquinoline quinone or salt thereof in preparing a medicament for preventing and treating prostatic hyperplasia and a medicinal composition.

The technical scheme is as follows: the invention provides application of pyrroloquinoline quinone or a salt thereof in preparing a medicament for preventing and treating prostatic hyperplasia.

Further, the pyrroloquinoline quinone or a salt thereof is administered by an enteral or parenteral or topical route.

Further, the administration route of the pyrroloquinoline quinone or the salt thereof is oral, sublingual, transdermal, intramuscular or subcutaneous, skin mucosa, vein and urethra.

A pharmaceutical composition for preventing and treating prostatic hyperplasia, which comprises pirrole quinoline quinone or its salt with preventing or treating effective amount.

Further, it contains 0.1-99.9% by weight of pyrroloquinoline quinone or a salt thereof as an active ingredient.

The pyrroloquinoline quinone or the salt thereof of the present invention can be obtained commercially and the purity thereof meets the pharmaceutical standard.

The pyrroloquinoline quinone or the salt thereof of the present invention can be used alone or in the form of a pharmaceutical composition. The pharmaceutical composition comprises the pyrroloquinoline quinone or the salt thereof of the present invention as an active ingredient and a pharmaceutically acceptable carrier. Wherein, the pharmaceutical composition of the invention contains 0.1 to 99.9 weight percent of the pyrroloquinoline quinone or the salt thereof as an active ingredient. The medicinal carrier does not damage the pharmaceutical activity of the pyrroloquinoline quinone or the salt thereof, and the effective dosage of the pyrroloquinoline quinone or the salt thereof is nontoxic to human bodies.

Such pharmaceutically acceptable carriers include, but are not limited to: lecithin, aluminum stearate, alumina, ion exchange materials, self-emulsifying drug delivery systems, tweens or other surfactants, serum proteins, buffer substances such as phosphates, glycine, sorbic acid, water, salts, electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, magnesium silicate, mixtures of saturated fatty acid partial glycerides, and the like.

Other conventional pharmaceutical adjuvants such as binder (e.g. microcrystalline cellulose), filler (e.g. starch, glucose, anhydrous lactose and lactose beads), disintegrant (e.g. crosslinked PVP, croscarmellose sodium, low-substituted hydroxypropylcellulose), lubricant (e.g. magnesium stearate), and absorption enhancer, adsorption carrier, flavoring agent, sweetening agent, excipient, diluent, wetting agent, etc.

The pyrroloquinoline quinone or salt thereof and the pharmaceutical composition thereof of the present invention can be prepared according to a conventional method in the art and administered by an enteral or parenteral or topical route. The oral preparation comprises capsule, tablet, oral liquid, granule, pill, powder, unguent, pellet, etc.; parenteral preparations include injections and the like; topical preparations include creams, patches, ointments, sprays, and the like.

The formulation form of the pyrroloquinoline quinone or the salt thereof and the pharmaceutical composition thereof of the present invention is preferably an oral formulation.

The administration route of the pyrroloquinoline quinone or the salt thereof and the pharmaceutical composition thereof of the present invention may be oral, sublingual, transdermal, intramuscular or subcutaneous, skin mucosa, vein, urethra, etc.

Has the advantages that: the pyrroloquinoline quinone or the pyrroloquinoline quinone salt has a good inhibition effect on prostatic hyperplasia, can obviously reduce the prostatic index, improve the SOD activity and reduce MDA (multidrug-associated disease) in serum and the like.

Drawings

FIG. 1 is a histopathological examination of prostate gland of a prostate-proliferating animal with different drugs, wherein A is a youth blank group, B is a youth model group, C is an elderly model group, D is a pyrroloquinoline quinone disodium salt high dose group, E is a pyrroloquinoline quinone disodium salt low dose group, and F is a finasteride group;

FIG. 2 is the immunohistochemical results for interleukin-1 a (IL-1a), wherein A is the youth blank group, B is the youth model group, C is the elderly model group, D is the pyrroloquinoline quinone disodium salt high dose group, E is the pyrroloquinoline quinone disodium salt low dose group, F is the finasteride group;

FIG. 3 is the results of Prostate Acid Phosphatase (PAP) immunohistochemistry in which group A young blank, group B young model, group C old model, group D pyrroloquinoline quinone disodium high dose, group E pyrroloquinoline quinone disodium low dose, group F finasteride.

Detailed Description

The pyrroloquinoline quinone disodium salt has the effect of preventing and treating prostatic hyperplasia caused by testosterone propionate.

1. Experimental Material

1.1 Experimental animals

SPF-ICR mice (aged group: 8 months old, body weight 40-57 g; young group: 8 weeks old, body weight 30-40g) were provided by the university of Nantong laboratory animal center, and the certification numbers: SCXK (Su) 2014-.

1.2 drugs and reagents

The positive drug finasteride (5 mg/tablet, Hangzhou Moshadong pharmaceutical Co., Ltd.) is administered in an amount of 1.67mg/kg, which is 20 times the clinical dose (5mg/60 kg).

Pyrroloquinoline quinone disodium salt (Sigma) was administered in an amount of 10mg/kg and 1 mg/kg.

Experimental feed: complete nutrition pellet feed (Nanjing collaborative animal feed factory).

Superoxide dismutase (SOD) kit and Malondialdehyde (MDA) kit (Nanjing institute of bioengineering) were assayed according to the instructions and techniques known to those skilled in the art.

1.3 Experimental statistics method

Statistical analysis was performed using SPSS13.0 statistical software, results were expressed as mean. + -. standard deviation (X. + -. S), analysis of variance was used, and LSD-t test was used for pairwise comparisons between groups. The difference is statistically significant when P is less than 0.05.

1.4 Experimental conditions

Room temperature 22 + -2 deg.C, humidity 55-65%, proper illumination, and good ventilation.

1.5 Equipment for experiments

FA1004 electronic balance, shanghaiengaku balance factory; LDZ5-2 type centrifuge, Beijing Eriki centrifuge, Inc.; ultraviolet spectrophotometer, general-purpose factory for Beijing Pujingyu.

2. Experimental methods

40-57g of aged ICR mice; young group ICR mice, 30-40 g: all are randomly grouped according to weight:

(1) young model group: 10 per group, and 10mL/kg of physiological saline is given;

(2) old age model group: 10 per group, and 10mL/kg of physiological saline is given;

(3) high dose group for elderly: 10 per group, 10 mg/kg;

(4) elderly dosing low dose group: 10 per group, 1 mg/kg;

(5) finasteride group: each group contained 10 individuals at 1.67 mg/kg.

The animals were gavaged (administration volume 10mL/kg), molding was started 2 weeks after administration, and all mice in each group were injected subcutaneously with testosterone propionate dissolved in olive oil 5mg/kg (25 mg/branch, 0.5mg/mL, 0.1 mL/mouse), and the blank group was injected with olive oil alone (0.1 mL/mouse). The injection is performed 1 time a day, and after 21 days of continuous injection, 2 testosterone propionate model groups are sacrificed, and the shape, the color, the texture and the adhesion condition with surrounding tissues of the prostate are observed in an anatomical mode. Weighing prostate and examining pathological examination by a section microscope to confirm whether the model is successfully made. The drug administration is continued for 30 days, and 24 hours after the last drug administration, fasting without water is performed, and pharmacodynamic evaluation is performed:

(1) the prostate tissue was dissected and isolated from each group of animals and weighed, and the organ index was calculated:

prostate Index (PI) ═ M₁/10×M₀(ii) a Wherein M is₁Prostate mean weight; m₀Mean body weight

(2) Serum was taken before sacrifice and the contents of SOD and MDA were determined using a full wavelength spectrophotometer.

(3) The prostate of each group of animals was taken for pathological examination and scored. The change of glandular epithelial cells, cytoplasm, nipple, glandular cavity endocrine, glandular connective tissue and the like among all the groups is comparatively observed.

(4) The method adopts immunohistochemistry method to determine interleukin 1a (IL-1a) and Prostatic Acid Phosphatase (PAP) in prostatic hyperplastic tissue after drug intervention.

3. Results of the experiment

3.1 Effect of pyrroloquinoline quinone disodium salt on Prostate Index (PI)

After different medicines are used for preventing and administrating for 2 weeks, testosterone propionate is continuously injected subcutaneously for 3 weeks to cause prostatic hyperplasia, at the end of an experiment, the weight of the prostate of an aged model group mouse is obviously increased, the prostate index is obviously increased, and the prostate weight is obviously different from that of a young blank group and a young model group (P is less than 0.01 and P is less than 0.05).

The weight of the prostate of the aged mice in the pyrroloquinoline quinone disodium salt administration group is not obviously increased, the prostate index is obviously different from the model group ratio (P is less than 0.01, and P is less than 0.05), and the drug effect is equivalent to that of the positive drug finasteride.

TABLE 1 comparison of experimental end body weight, prostate weight and prostate index for each group of animals

3.2 Effect of pyrroloquinoline quinone disodium salt on SOD and MDA in serum of animals with prostatic hyperplasia

After different drugs are prevented from being administered for 2 weeks, testosterone propionate is continuously injected subcutaneously for 3 weeks to cause prostatic hyperplasia, at the end of an experiment, animal serum is taken to detect SOD activity and MDA content according to a kit method, the SOD activity in the serum of mice in an old model group and a young model group is obviously reduced, and the SOD activity is obviously different from that in the young blank group (P is less than 0.01); the serum MDA content of mice in the old model group and the young model group is obviously increased, and has significant difference (P is less than 0.01) compared with the young blank group;

after the pyrroloquinoline quinone disodium salt is administrated, the SOD activity in the blood serum of a mouse is obviously improved, and the SOD activity is obviously different from that of an old model group and a young model group (P is less than 0.01). The SOD activity in the blood serum of the finasteride mouse is increased, but the statistic significance is not shown compared with the old model group and the young model group. After the pyrroloquinoline quinone disodium salt is administrated, the MDA content in the blood serum of a mouse is obviously reduced, and the mouse has a significant difference (P is less than 0.01) compared with an old model group and a young model group.

TABLE 2 comparison of SOD activity and MDA concentration in plasma of various groups of animals

3.3 pathological histology of prostate (FIG. 1)

Comparison with young blank (normal prostate tissue) revealed: the mouse of the old model group has different prostate gland sizes, local gland epithelial cells obviously proliferate, the gland epithelia are in a high column shape, folds form papillary bulges with different sizes into the gland cavities, and interstitial muscle connective tissues obviously proliferate. Pyrroloquinoline quinone disodium salt high dose group (8): compared with the model group, the hyperplasia of glandular epithelium is reduced, partial glands are recovered to be normal, the cavities are filled with eosin-stained secretion, the hyperplasia degree of interstitial muscle connective tissues is also reduced, and a little inflammatory cell infiltration exists in individual interstitium. Pyrroloquinoline quinone disodium salt low dose group (7): compared with the aged model group, the hyperplasia of glandular epithelium is slightly reduced, small papillary protrusions are formed by the glandular mucosal folds of 1 case, partial glands are slightly expanded locally, interstitial sarcoid tissues are slightly proliferated, and a small amount of inflammatory cell infiltration exists in individual interstitium. Finasteride group (8): compared with the model group, the hyperplasia of glandular epithelium is obviously reduced, the connective tissue of interstitial muscle is slightly hyperplastic, the interstitial tissue has no inflammatory cell infiltration phenomenon, 1 interstitial tissue has bleeding phenomenon, and 1 interstitial tissue around gland has diffuse inflammatory cell infiltration.

And (4) conclusion: the main pathological changes of the prostate of the model mouse are that the glandular epithelial cells of the local prostate gland of the mouse obviously proliferate, mucosal folds form papillary bulges with different sizes into the glandular cavity, the glandular epithelium partially shed and a small amount of secretion can be seen in the glandular cavity, interstitial myoconnective tissues obviously proliferate, blood vessels expand and hyperemia, and more inflammatory cell infiltration can be seen in the interstitial tissues around the proliferated glands of individual mice, and the individual mice mainly comprise lymphocytes and macrophages. Prostate tissue of aged mice also proliferated to different degrees. The overall effect is better by a pyrroloquinoline quinone disodium salt high dose group, a pyrroloquinoline quinone disodium salt low dose group and a finasteride medicine group.

3.4 Effect of pyrroloquinoline quinone disodium salt on the expression of IL-1a and PAP in prostate tissue

The results of interleukin-1 a (IL-1a) immunohistochemistry (FIG. 2) show: prostate tissue from the young blank showed negative expression, and IL-1a showed negative (-) expression. Young model group: prostate tissue appears brownish yellow and IL-1a appears strongly positive (++) expression. Old age model group: prostate tissue appears brownish yellow and IL-1a appears strongly positive (++) expression. Pyrroloquinoline quinone disodium salt high dose group: prostate tissue was negative and IL-1a was weakly positive (+) expressed. Pyrroloquinoline quinone disodium salt low dose group: prostate tissue appears yellowish and IL-1a appears weakly positive (+) expression. Finasteride group: prostate tissue appears yellowish and IL-1a appears weakly positive (+) expression.

The results of Prostate Acid Phosphatase (PAP) immunohistochemistry (fig. 3) show: young blank group: prostate tissue appears negative and PAP appears negative (-) expression. Young model group: prostate tissue appears brownish yellow and PAP appears strongly positive (++) expressed. Old age model group: prostate tissue appears brownish yellow and PAP appears strongly positive (++) expressed. Pyrroloquinoline quinone disodium salt high dose group: prostate tissue was negative and IL-1a was weakly positive (+) expressed. Pyrroloquinoline quinone disodium salt low dose group: prostate tissue appears yellowish and IL-1a appears weakly positive (+) expression. Finasteride group: prostate tissue appears yellowish and IL-1a appears weakly positive (+) expression.