COMBINATION OF COMPOUNDS FOR TREATING VASCULAR DISEASE COMPRISING A PDE5 INHIBITOR, ARGININE, AND N-ACETYLCYSTEINE - Patent application

Abstract

The present disclosure relates to the technical field of medicine.In particular, the present disclosure provides a pharmaceutical combination / composition comprising a phosphodiesterase type 5 (PD1 5) inhibitor, arginine and N-acetylcysteine, and its application in the treatment of cardiovascular disease and erectile dysfunction.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of Taiwan (ROC) Patent Application No. 111104639, filed February 9, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] Field The present disclosure relates to the technical field of medicine. In particular, the present disclosure provides a pharmaceutical combination / composition comprising a phosphodiesterase type 5 (PDE5) inhibitor, arginine, and N-acetylcysteine, and its application in the treatment of cardiovascular disease and erectile dysfunction. [Background technology]

[0003] Cardiovascular disease is a very serious health problem in modern society. In addition to a high-salt and high-fat diet, with industrial development, the possibility of cardiovascular damage is increasing year by year due to the inhalation of various environmental pollutants or compounds in daily life.

[0004] Erectile dysfunction (ED) is a problem that has received significant attention in recent years. In addition to psychological factors, erectile dysfunction may be caused by abnormal bodily function. A significant amount of research has been conducted into the use of medications to treat and alleviate ED.

[0005] Phosphodiesterase type 5 inhibitors (PDE5 inhibitors) are vasodilators that act by blocking the degradative action of cGMP-specific phosphodiesterase type 5 (PDE5) on cyclic GMP in smooth muscle cells lining blood vessels supplying various tissues. Part of the physiological process of vasodilation involves the release of nitric oxide (NO) by vascular endothelial cells, which then diffuses to nearby vascular smooth muscle cells. There, NO activates soluble guanylate cyclase, which converts guanosine triphosphate (GTP) to cyclic guanosine monophosphate (cGMP), the main effector of this system. PDE5 inhibitors prolong the action of cGMP by preventing its breakdown by the enzyme PDE5, which is found throughout the body (Goldstein I, Lue TF, Padma-Nathan H, Rosen RC, Steers WD, Wicker PA (May 1998). "Oral sildenafil in the treatment of erectile dysfunction. Sildenafil Study Group," The New England Journal of Medicine. 338 (20): 1397-404). PDE5 inhibitors are contraindicated in combination with alpha-blockers, soluble guanylate cyclase stimulators, or nitrates, as this may cause excessively low blood pressure. PDE5 inhibitors may cause headache, abdominal pain, diarrhea, flushing, dizziness, weakness, pruritus, and erythema after administration. Summary of the Invention [Problem to be solved by the invention]

[0006] To date, there remains a need to develop safer, more effective pharmaceutical compositions or combinations useful for treating or preventing cardiovascular disease and ED. [Means for solving the problem]

[0007] The present disclosure relates to a PDE5 inhibitor or its pharmaceutically acceptable salt in combination with arginine and / or N-acetylcysteine ​​(NAC), which surprisingly increases the reaction rate of the PDE5 inhibitor.In particular, the reaction time is shortened and endogenous free radicals generated by rapid vasodilation are further removed, which can protect blood vessels.

[0008] In one embodiment, the present disclosure relates to a pharmaceutical combination or composition comprising a phosphodiesterase type 5 (PDE5) inhibitor or a pharmaceutically acceptable salt thereof, arginine, and N-acetylcysteine.

[0009] In one embodiment, the present disclosure relates to a pharmaceutical composition comprising a PDE5 inhibitor or a pharmaceutically acceptable salt thereof, arginine, N-acetylcysteine, and a pharmaceutically acceptable excipient.

[0010] In one embodiment, the PDE5 inhibitor of the present disclosure is selected from sildenafil, tadalafil, and vardenafil, and / or pharmaceutically acceptable salts thereof.

[0011] In one embodiment, the amount of PDE5 inhibitor in the pharmaceutical combination or composition described herein ranges from about 0.1% (w / w) to about 50% (w / w). Specific embodiments of the amount of PDE5 inhibitor include, but are not limited to, about 0.1% (w / w) to about 45% (w / w), about 0.1% (w / w) to about 40% (w / w), about 0.1% (w / w) to about 35% (w / w), about 0.1% (w / w) to about 30% (w / w), about 0.1% (w / w) to about 25% (w / w), about 0.1% (w / w) to about 20% (w / w), about 0.1% (w / w) to about 15% (w / w), and the like. w / w), about 0.1% (w / w) to about 10% (w / w), about 0.1% (w / w) to about 5% (w / w), about 0.1% (w / w) to about 2.5% (w / w), about 0.1% (w / w) to about 1% (w / w), about 0.5% (w / w) to about 50% (w / w), about 0.5% (w / w) to about 45% (w / w), about 0.5% (w / w) to about 40% (w / w), about 0.5% (w / w) to about 35% (w / w), about 0.5% (w / w) to about 30% (w / w), about 0.5% (w / w) to about 25% (w / w), about 0.5% (w / w) to about 20% (w / w), about 0.5% (w / w) to about 15% (w / w), about 0.5% (w / w) to about 10% (w / w), about 0.5% (w / w) to about 5% (w / w), about 0.5% (w / w) to about 2.5% (w / w), about 0.5% (w / w) to about 1% (w / w), about 1.0% (w / w) to about 50% (w / w), about 1.0% (w / w) to about 45% (w / w), about 1.0% (w / w) to about 40% (w / w), about 1.0% (w / w) to about 35% (w / w), about 1.0% (w / w) to about 30% (w / w), about 1.0% (w / w) to about 25% (w / w), about 1.0% (w / w) to about 20% (w / w), about 1.0% (w / w) to about 15% (w / w), about 1.0% (w / w) to about 10% (w / w), about 1.0% (w / w) to about 5% (w / w), about 5% (w / w) to about 50% (w / w), about 5% (w / w) to about 45% (w / w), about 5% (w / w) to about 40% (w / w), about 5% (w / w) to about 35% (w / w), about 5% (w / w) to about 30% (w / w), about 5% (w / w) to about 25% (w / w), about 5% (w / w) to about 20% (w / w), about 5% (w / w) to about 15% (w / w), about 5% (w / w) to about 10% (w / w), about 10% (w / w) to Approximately 50% (w / w), approximately 10% (w / w) to approximately 45% (w / w), approximately 10% (w / w) to approximately 40% (w / w), approximately 10% (w / w) to approximately 35% (w / w), approximately 10% (w / w) to approximately 30% (w / w), approximately 10% (w / w) to approximately 25% (w / w), approximately 10% (w / w) to approximately 20% (w / w), approximately 15% (w / w) to approximately 50% (w / w), approximately 15% (w / w) to approximately 45% (w / w), approximately 15% (w / w) to approximately 40% (w / w), approximately 15% (w / w) ~ about 35% (w / w), about 15% (w / w) to about 30% (w / w), about 15% (w / w) to about 25% (w / w), about 20% (w / w) to about 50% (w / w), about 20% (w / w) to about 45% (w / w), about 20% (w / w) to about 40% (w / w), about 20% (w / w) to about 35% (w / w), about 20% (w / w) to about 30% (w / w), about 20% (w / w) to about 25% (w / w), about 25% (w / w) to about 50% (w / w), about 25% (w / w) ) to about 45% (w / w), about 25% (w / w) to about 40% (w / w), about 25% (w / w) to about 35% (w / w), about 30% (w / w) to about 50% (w / w), about 30% (w / w) to about 45% (w / w), about 30% (w / w) to about 40% (w / w), about 35% (w / w) to about 50% (w / w), about 35% (w / w) to about 45% (w / w), about 40% (w / w) to about 50% (w / w), and about 40% (w / w) to about 45% (w / w).

[0012] In one embodiment, the amount of arginine in a pharmaceutical combination or composition described herein ranges from about 4.0% (w / w) to about 80% (w / w). Specific embodiments of the amount of arginine include, but are not limited to, about 4.0% (w / w) to about 75% (w / w), about 4.0% (w / w) to about 70% (w / w), about 4.0% (w / w) to about 65% (w / w), about 4.0% (w / w) to about 60% (w / w), about 4.0% (w / w) to about 55% (w / w), about 4.0% (w / w) to about 50% (w / w), about 4.0% (w / w) to about 45% (w / w), about 4.0% (w / w) to about 40% (w / w), about 4.0% (w / w) to about 35% (w / w), and about 4.0% (w / w). ) to about 30% (w / w), about 4.0% (w / w) to about 25% (w / w), about 4.0% (w / w) to about 20% (w / w), about 4.0% (w / w) to about 15% (w / w), about 4.0% (w / w) to about 10% (w / w), about 10.0% (w / w) to about 80% (w / w), about 10.0% (w / w) to about 75% (w / w), about 10.0% (w / w) to about 70% (w / w), about 10.0% (w / w) to about 65% (w / w), about 10.0% (w / w) to about 60% (w / w), about 10.0% (w / w) to about 55% (w / w), about 10 0.0% (w / w) to about 50% (w / w), about 10.0% (w / w) to about 45% (w / w), about 10.0% (w / w) to about 40% (w / w), about 10.0% (w / w) to about 35% (w / w), about 10.0% (w / w) to about 30% (w / w), about 10.0% (w / w) to about 25% (w / w), about 10.0% (w / w) to about 20% (w / w), about 10.0% (w / w) to about 15% (w / w), about 15.0% (w / w) to about 80% (w / w), about 15.0% (w / w) to about 75% (w / w), about 15.0% (w / w) to about 7 0% (w / w), about 15.0% (w / w) to about 65% (w / w), about 15.0% (w / w) to about 60% (w / w), about 15.0% (w / w) to about 55% (w / w), about 15.0% (w / w) to about 50% (w / w), about 15.0% (w / w) to about 45% (w / w), about 15.0% (w / w) to about 40% (w / w), about 15.0% (w / w) to about 35% (w / w), about 15.0% (w / w) to about 30% (w / w), about 15.0% (w / w) to about 25% (w / w), about 15.0% (w / w) to about 20% (w / w), about 20.0% (w / w) to approximately 80% (w / w), approximately 20.0% (w / w) to approximately 75% (w / w), approximately 20.0% (w / w) to approximately 70% (w / w), approximately 20.0% (w / w) to approximately 65% ​​(w / w), approximately 20.0% (w / w) to approximately 60% (w / w), approximately 20.0% (w / w) to approximately 55% ( w / w), about 20.0% (w / w) to about 50% (w / w), about 20.0% (w / w) to about 45% (w / w), about 20.0% (w / w) to about 40% (w / w), about 20.0% (w / w) to about 35% (w / w), about 20.0% (w / w) to about 30% (w / w), about 20.0% (w / (w / w) to about 25% (w / w), about 25.0% (w / w) to about 80% (w / w), about 25.0% (w / w) to about 75% (w / w), about 25.0% (w / w) to about 70% (w / w), about 25.0% (w / w) to about 65% (w / w), about 25.0% (w / w) to about 60% (w / w), about 25.0% (w / w) to approximately 55% (w / w), approximately 25.0% (w / w) to approximately 50% (w / w), approximately 25.0% (w / w) to approximately 45% (w / w), approximately 25.0% (w / w) to approximately 40% (w / w), approximately 25.0% (w / w) to approximately 35% (w / w), approximately 25.0% (w / w) to approximately 30 % (w / w), approximately 30.0% (w / w) to approximately 80% (w / w), approximately 30.0% (w / w) to approximately 75% (w / w), approximately 30.0% (w / w) to approximately 70% (w / w), approximately 30.0% (w / w) to approximately 65% ​​(w / w), approximately 30.0% (w / w) to approximately 60% (w / w), approximately 30.0% (w / w) to about 55% (w / w), about 30.0% (w / w) to about 50% (w / w), about 30.0% (w / w) to about 45% (w / w), about 30.0% (w / w) to about 40% (w / w), about 30.0% (w / w) to about 35% (w / w), about 35.0% (w / w) to about 80% (w / w) ), about 35.0% (w / w) to about 75% (w / w), about 35.0% (w / w) to about 70% (w / w), about 35.0% (w / w) to about 65% (w / w), about 35.0% (w / w) to about 60% (w / w), about 35.0% (w / w) to about 55% (w / w), about 35.0% (w / w) to Approximately 50% (w / w), approximately 35.0% (w / w) to approximately 45% (w / w), approximately 35.0% (w / w) to approximately 40% (w / w), approximately 40.0% (w / w) to approximately 80% (w / w), approximately 40.0% (w / w) to approximately 75% (w / w), approximately 40.0% (w / w) to approximately 70% (w / w), approximately 40.0% (w / w) to about 65% (w / w), about 40.0% (w / w) to about 60% (w / w), about 40.0% (w / w) to about 55% (w / w), about 40.0% (w / w) to about 50% (w / w), about 40.0% (w / w) to about 45% (w / w), about 45.0% (w / w) to about 80% (w / w), about 45.0% (w / w) to about 75% (w / w), about 45.0% (w / w) to about 70% (w / w), about 45.0% (w / w) ) to about 65% (w / w), about 45.0% (w / w) to about 60% (w / w), about 45.0% (w / w) to about 55% (w / w), about 45.0% (w / w) to about 50% (w / w), about 50.0% (w / w) to about 80% (w / w), about 50.0% (w / w) to about 75% (w / w), about 50.0% (w / w) to about 70% (w / w), about 50.0% (w / w) to about 65% (w / w), about 50.0% (w / w) to about 60% ( w / w), about 50.0% (w / w) to about 55% (w / w), about 55.0% (w / w) to about 80% (w / w), about 55.0% (w / w) to about 75% (w / w), about 55.0% (w / w) to about 70% (w / w), about 55.0% (w / w) to about 65% (w / w), about 55.0% (w / w) to about 60% (w / w), about 60.0% (w / w) to about 80% (w / w), about 60.0% (w / w) to about 75% (w / w), about Examples include 60.0% (w / w) to about 70% (w / w), about 60.0% (w / w) to about 65% (w / w), about 65.0% (w / w) to about 80% (w / w), about 65.0% (w / w) to about 75% (w / w), about 65.0% (w / w) to about 70% (w / w), about 70.0% (w / w) to about 80% (w / w), about 70.0% (w / w) to about 75% (w / w), or about 75.0% (w / w) to about 80% (w / w).

[0013] In one embodiment, the amount of N-acetylcysteine ​​in the pharmaceutical combination or composition described herein ranges from about 4% (w / w) to about 80% (w / w). Specific embodiments of the amount of arginine include, but are not limited to, about 4.0% (w / w) to about 75% (w / w), about 4.0% (w / w) to about 70% (w / w), about 4.0% (w / w) to about 65% (w / w), about 4.0% (w / w) to about 60% (w / w), about 4.0% (w / w) to about 55% (w / w), about 4.0% (w / w) to about 50% (w / w), about 4.0% (w / w) to about 45% (w / w), about 4.0% (w / w) to about 40% (w / w), about 4.0% (w / w) to about 35% (w / w), and about 4.0% (w / w). ) to about 30% (w / w), about 4.0% (w / w) to about 25% (w / w), about 4.0% (w / w) to about 20% (w / w), about 4.0% (w / w) to about 15% (w / w), about 4.0% (w / w) to about 10% (w / w), about 10.0% (w / w) to about 80% (w / w), about 10.0% (w / w) to about 75% (w / w), about 10.0% (w / w) to about 70% (w / w), about 10.0% (w / w) to about 65% (w / w), about 10.0% (w / w) to about 60% (w / w), about 10.0% (w / w) to about 55% (w / w), about 10 0.0% (w / w) to about 50% (w / w), about 10.0% (w / w) to about 45% (w / w), about 10.0% (w / w) to about 40% (w / w), about 10.0% (w / w) to about 35% (w / w), about 10.0% (w / w) to about 30% (w / w), about 10.0% (w / w) to about 25% (w / w), about 10.0% (w / w) to about 20% (w / w), about 10.0% (w / w) to about 15% (w / w), about 15.0% (w / w) to about 80% (w / w), about 15.0% (w / w) to about 75% (w / w), about 15.0% (w / w) to about 7 0% (w / w), about 15.0% (w / w) to about 65% (w / w), about 15.0% (w / w) to about 60% (w / w), about 15.0% (w / w) to about 55% (w / w), about 15.0% (w / w) to about 50% (w / w), about 15.0% (w / w) to about 45% (w / w), about 15.0% (w / w) to about 40% (w / w), about 15.0% (w / w) to about 35% (w / w), about 15.0% (w / w) to about 30% (w / w), about 15.0% (w / w) to about 25% (w / w), about 15.0% (w / w) to about 20% (w / w), about 20.0% (w / w) to approximately 80% (w / w), approximately 20.0% (w / w) to approximately 75% (w / w), approximately 20.0% (w / w) to approximately 70% (w / w), approximately 20.0% (w / w) to approximately 65% ​​(w / w), approximately 20.0% (w / w) to approximately 60% (w / w), approximately 20.0% (w / w) to approximately 55% ( w / w), about 20.0% (w / w) to about 50% (w / w), about 20.0% (w / w) to about 45% (w / w), about 20.0% (w / w) to about 40% (w / w), about 20.0% (w / w) to about 35% (w / w), about 20.0% (w / w) to about 30% (w / w), about 20.0% (w / (w / w) to about 25% (w / w), about 25.0% (w / w) to about 80% (w / w), about 25.0% (w / w) to about 75% (w / w), about 25.0% (w / w) to about 70% (w / w), about 25.0% (w / w) to about 65% (w / w), about 25.0% (w / w) to about 60% (w / w), about 25.0% (w / w) to approximately 55% (w / w), approximately 25.0% (w / w) to approximately 50% (w / w), approximately 25.0% (w / w) to approximately 45% (w / w), approximately 25.0% (w / w) to approximately 40% (w / w), approximately 25.0% (w / w) to approximately 35% (w / w), approximately 25.0% (w / w) to approximately 30 % (w / w), approximately 30.0% (w / w) to approximately 80% (w / w), approximately 30.0% (w / w) to approximately 75% (w / w), approximately 30.0% (w / w) to approximately 70% (w / w), approximately 30.0% (w / w) to approximately 65% ​​(w / w), approximately 30.0% (w / w) to approximately 60% (w / w), approximately 30.0% (w / w) to about 55% (w / w), about 30.0% (w / w) to about 50% (w / w), about 30.0% (w / w) to about 45% (w / w), about 30.0% (w / w) to about 40% (w / w), about 30.0% (w / w) to about 35% (w / w), about 35.0% (w / w) to about 80% (w / w) ), about 35.0% (w / w) to about 75% (w / w), about 35.0% (w / w) to about 70% (w / w), about 35.0% (w / w) to about 65% (w / w), about 35.0% (w / w) to about 60% (w / w), about 35.0% (w / w) to about 55% (w / w), about 35.0% (w / w) to Approximately 50% (w / w), approximately 35.0% (w / w) to approximately 45% (w / w), approximately 35.0% (w / w) to approximately 40% (w / w), approximately 40.0% (w / w) to approximately 80% (w / w), approximately 40.0% (w / w) to approximately 75% (w / w), approximately 40.0% (w / w) to approximately 70% (w / w), approximately 40.0% (w / w) to about 65% (w / w), about 40.0% (w / w) to about 60% (w / w), about 40.0% (w / w) to about 55% (w / w), about 40.0% (w / w) to about 50% (w / w), about 40.0% (w / w) to about 45% (w / w), about 45.0% (w / w) to about 80% (w / w), about 45.0% (w / w) to about 75% (w / w), about 45.0% (w / w) to about 70% (w / w), about 45.0% (w / w) ) to about 65% (w / w), about 45.0% (w / w) to about 60% (w / w), about 45.0% (w / w) to about 55% (w / w), about 45.0% (w / w) to about 50% (w / w), about 50.0% (w / w) to about 80% (w / w), about 50.0% (w / w) to about 75% (w / w), about 50.0% (w / w) to about 70% (w / w), about 50.0% (w / w) to about 65% (w / w), about 50.0% (w / w) to about 60% ( w / w), about 50.0% (w / w) to about 55% (w / w), about 55.0% (w / w) to about 80% (w / w), about 55.0% (w / w) to about 75% (w / w), about 55.0% (w / w) to about 70% (w / w), about 55.0% (w / w) to about 65% (w / w), about 55.0% (w / w) to about 60% (w / w), about 60.0% (w / w) to about 80% (w / w), about 60.0% (w / w) to about 75% (w / w), about Examples include 60.0% (w / w) to about 70% (w / w), about 60.0% (w / w) to about 65% (w / w), about 65.0% (w / w) to about 80% (w / w), about 65.0% (w / w) to about 75% (w / w), about 65.0% (w / w) to about 70% (w / w), about 70.0% (w / w) to about 80% (w / w), about 70.0% (w / w) to about 75% (w / w), or about 75.0% (w / w) to about 80% (w / w).

[0014] In one embodiment, the amounts of PDE5 inhibitor, arginine, and N-acetylcysteine ​​range from about 0.1% (w / w) to about 50% (w / w), from about 4.0% (w / w) to about 80% (w / w), and from about 4% (w / w) to about 80% (w / w), respectively. Certain embodiments of the amounts of PDE5 inhibitor, arginine, and N-acetylcysteine ​​are within the ranges described herein.

[0015] In one embodiment, the amount of PDE5 inhibitor in the pharmaceutical combination or composition described herein ranges from about 0.5 mg to about 250 mg. Specific embodiments of the amount of PDE5 inhibitor include, but are not limited to, about 0.5 mg to about 250 mg, about 0.5 mg to about 240 mg, about 0.5 mg to about 230 mg, about 0.5 mg to about 220 mg, 0.5 mg to about 210 mg, about 0.5 mg to about 200 mg, about 0.5 mg to about 190 mg, about 0.5 mg to about 180 mg, about 0.5 mg to about 170 mg, about 0.5 mg to about 160 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 140 mg, about 0.5 mg to about 130 mg, about 0.5 mg to about 120 mg, and about 0.5 mg to about 160 mg. mg ~ about 110mg, about 0.5mg - about 100mg, about 0.5mg - about 90mg, about 0.5mg - about 80mg, about 0.5mg - about 70mg, about 0.5mg - about 60mg, about 0.5mg - about 50mg, about 0.5mg - about 40mg, about 0.5mg - about 30mg, about 0 .5mg to about 20mg, about 0.5mg to about 10mg, about 0.5mg to about 5mg, about 0.5mg to about 1mg, about 1mg to about 250mg, about 1mg to about 240mg, about 1mg to about 230mg, about 1mg to about 220mg, about 1mg to about 210mg, about 1mg to about 200 mg, about 1mg to about 190mg, about 1mg to about 180mg, about 1mg to about 170mg, about 1mg to about 160mg, about 1mg to about 150mg, about 1mg to about 140mg, about 1mg to about 130mg, about 1mg to about 120mg, about 1mg to about 110mg, about 1mg to about 100mg, about 1mg to about 90mg, about 1mg to about 80mg, about 1mg to about 70mg, about 1mg to about 60mg, about 1mg to about 50mg, about 1mg to about 40mg, about 1mg to about 30mg, about 1mg to about 20mg, about 1mg to about 10mg, about 1mg to about 5mg, about 5 mg~250mg, 5mg~240mg, 5mg~230mg, 5mg~220mg, 5mg~210mg, 5mg~200mg, 5mg~190mg, 5mg~180mg, 5mg~170mg, 5mg~160mg , about 5 mg to about 150 mg, about 5 mg to about 140 mg, about 5 mg to about 130 mg, about 5 mg to about 120 mg, about 5 mg to about 110 mg, about 5 mg to about 100 mg, about 5 mg to about 90 mg, about 5 mg to about 80 mg, about 5 mg to about 70 mg, about 5 mg to about 60 mg,about 5 mg to about 50 mg, about 5 mg to about 40 mg, about 5 mg to about 30 mg, about 5 mg to about 20 mg, about 5 mg to about 10 mg, about 10 mg to about 250 mg, about 10 mg to about 240 mg, about 10 mg to about 230 mg, about 10 mg to about 220 mg, about 10 mg to about 210 mg, about 10 mg to about 200 mg, about 10 mg to about 190 mg, about 10 mg to about 180 mg, about 10 mg to about 170 mg, about 10 mg to about 160 mg, about 10 mg to about 150 mg, about 10 mg to about 140 mg, about 10 mg to about 130 mg, about 10 mg to about 120 mg, about 10 mg to about 110 mg g, about 10 mg to about 100 mg, about 10 mg to about 90 mg, about 10 mg to about 80 mg, about 10 mg to about 70 mg, about 10 mg to about 60 mg, about 10 mg to about 50 mg, about 10 mg to about 40 mg, about 10 mg to about 30 mg, about 10 mg to about 20 mg, about 20 mg to about 250 mg, about 20 mg to about 240 mg, about 20 mg to about 230 mg, about 20 mg to about 220 mg, about 20 mg to about 210 mg, about 20 mg to about 200 mg, about 20 mg to about 190 mg, about 20 mg to about 180 mg, about 20 mg to about 170 mg, about 20 mg to about 160 mg, about 20 mg to about 150 mg, about 20 mg to about 140 mg, about 20 mg to about 130 mg, about 20 mg to about 120 mg, about 20 mg to about 110 mg, about 20 mg to about 100 mg, about 20 mg to about 90 mg, about 20 mg to about 80 mg, about 20 mg to about 70 mg, about 20 mg to about 60 mg, about 20 mg to about 50 mg, about 20 mg to about 40 mg, about 20 mg to about 30 mg, about 30 mg to about 250 mg, 30 mg to about 240 mg, 30 mg to about 230 mg, 30 mg to about 220 mg, 30 mg to about 210 mg, about 30 mg to about 200 mg, about 30 mg to about 190 mg, about 30 mg to about 180 mg, about 30 mg to about 170 mg, about 30 mg to about 160 mg, about 30 mg to about 150 mg, about 30 mg to about 140 mg, about 30 mg to about 130 mg, about 30 mg to about 120 mg, about 30 mg to about 110 mg, about 30 mg to about 100 mg, about 30 mg to about 90 mg, about 30 mg to about 80 mg, about 30 mg to about 70 mg, about 30 mg to about 60 mg, about 30 mg to about 50 mg, about 30 mg to about 40 mg, about 40 mg to about 250 mg, about 40 mg to about 240 mg, about 40 mg to about 230 mg, about 40 mg to about 220 mg, about 40 mg to about 210 mg,about 40 mg to about 200 mg, about 40 mg to about 190 mg, about 40 mg to about 180 mg, about 40 mg to about 170 mg, about 40 mg to about 160 mg, about 40 mg to about 150 mg, about 40 mg to about 140 mg, about 40 mg to about 130 mg, about 40 mg to about 120 mg, about 40 mg to about 110 mg, about 40 mg to about 100 mg, about 40 mg to about 90 mg, about 40 mg to about 80 mg, about 40 mg to about 70 mg, about 40 mg to about 60 mg, about 40 mg to about 50 mg, about 50 mg to about 250 mg, about 50 mg to about 240 mg, about 50 mg to about 230 mg, about 50 mg to about 220 mg, about 50 mg to about 210 mg, about 50 mg to about 200 mg, about 50 mg to about 190 mg, about 50 mg to about 180 mg, about 50 mg to about 170 mg, about 50 mg to about 160 mg, about 50 mg to about 150 mg, about 50 mg to about 140 mg, about 50 mg to about 130 mg, about 50 mg to about 120 mg, about 50 mg to about 110 mg, about 50 mg to about 100 mg, about 50 mg to about 90 mg, about 50 mg to about 80 mg, about 50 mg to about 70 mg, about 50 mg to about 60 mg, about 60 mg to about 250 mg, about 60 mg to about 240 mg, about 60 mg to about 230 mg, about 60 mg to about 220mg, about 60mg to about 210mg, about 60mg to about 200mg, about 60mg to about 190mg, about 60mg to about 180mg, about 60mg to about 170mg, about 60mg to about 160mg, about 60mg to about 150mg, about 60mg to about 140mg, about 60mg to about 130mg, about 60mg to about 120mg, about 60mg to about 110mg, about 60mg to about 100mg, about 60mg to about 90mg, about 60mg to about 80mg, about 60mg to about 70mg, about 70mg to about 250mg, about 70mg to about 240mg, about 70mg to about 230mg, about 70mg to about 220mg, about 7 0mg to about 210mg, about 70mg to about 200mg, about 70mg to about 190mg, about 70mg to about 180mg, about 70mg to about 170mg, about 70mg to about 160mg, about 70mg to about 150mg, about 70mg to about 140mg, about 70mg to about 130mg, about 70mg to about 120mg, about 70mg to about 110mg, about 70mg to about 100mg, about 70mg to about 90mg, about 70mg to about 80mg, about 80mg to about 250mg, about 80mg to about 240mg, about 80mg to about 230mg, about 80mg to about 220mg, about 80mg to about 210mg,about 80 mg to about 200 mg, about 80 mg to about 190 mg, about 80 mg to about 180 mg, about 80 mg to about 170 mg, about 80 mg to about 160 mg, about 80 mg to about 150 mg, about 80 mg to about 140 mg, about 80 mg to about 130 mg, about 80 mg to about 120 mg, about 80 mg to about 110 mg, about 80 mg to about 100 mg, about 80 mg to about 90 mg, about 90 mg to about 250 mg, 90 mg to about 240 mg, 90 mg to about 230 mg, 90 mg to about 220 mg, 90 mg to about 210 mg, 90 mg to about 200 mg, 90 mg to about about 190 mg, 90 mg to about 180 mg, about 90 mg to about 170 mg, about 90 mg to about 160 mg, about 90 mg to about 150 mg, about 90 mg to about 140 mg, about 90 mg to about 130 mg, about 90 mg to about 120 mg, about 90 mg to about 110 mg, about 90 mg to about 100 mg, about 100 mg to about 250 mg, about 100 mg to about 240 mg, about 100 mg to about 230 mg, about 100 mg to about 220 mg, about 100 mg to about 210 mg, about 100 mg to about 200 mg, about 100 mg to about 190 mg, about 100 mg to about 180 mg, about 100 mg to about 170 mg, about 100 mg to about 160 mg, about 100 mg to about 150 mg, about 100 mg to about 140 mg, about 100 mg to about 130 mg, about 100 mg to about 120 mg, about 100 mg to about 110 mg, about 110 mg to about 250 mg, about 110 mg to about 240 mg, about 110 mg to about 230 mg, about 110 mg to about 220 mg, about 110 mg to about 210 mg, about 110 mg to about 200 mg, about 110 mg to about 190 mg, about 110 mg to about 180 mg, about 110 mg to about 170 mg, about 110 mg to about 160 mg, about 110 mg to about 150 mg, about 110 mg to about 140 mg, about 110 mg to about 1 30mg, about 110mg to about 120mg, about 120mg to about 250mg, about 120mg to about 240mg, about 120mg to about 230mg, about 120mg to about 220mg, about 120mg to about 210mg, about 120mg to about 200mg, about 120mg to about 190mg, about 120mg to about 180mg, about 120mg to about 170mg, about 120mg to about 160mg, about 120mg to about 150mg, about 120mg to about 140mg, about 120mg to about 130mg, about 130mg to about 250mg, 130mg to about 240mg, about 130mg to about 230mg, about 130mg to about 220mg,About 130mg to about 210mg, about 130mg to about 200mg, about 130mg to about 190mg, about 130mg to about 180mg, about 130mg to about 170mg, about 130mg to about 160mg, about 130mg to about 150mg, about 130mg to about 140mg, about 130mg to about 250mg, about 130mg to about 240mg, about 130mg to about 230mg, about 130mg to about 220mg, about 130mg to about 210mg, about 130mg to about 200mg, about 130mg to about 190mg, about 130mg to about 180mg, about 130mg to about 170mg, about 130m g ~ about 160mg, about 130mg - about 150mg, about 130mg - about 140mg, about 140mg - about 250mg, about 140mg - about 240mg, about 140mg - about 230mg, about 14 0mg to about 220mg, about 140mg to about 210mg, about 140mg to about 200mg, about 140mg to about 190mg, about 140mg to about 180mg, about 140mg to about 170mg, Approximately 140mg to approximately 160mg, approximately 140mg to approximately 150mg, approximately 150mg to approximately 250mg, approximately 150mg to approximately 240mg, approximately 150mg to approximately 230mg, approximately 150mg to approximately 220 mg, about 150 mg to about 210 mg, about 150 mg to about 200 mg, about 150 mg to about 190 mg, about 150 mg to about 180 mg, about 150 mg to about 170 mg, about 150 mg to about 160 mg, about 200 mg to about 250 mg, about 200 mg to about 240 mg, about 200 mg to about 230 mg, about 200 mg to about 220 mg, and about 200 mg to about 210 mg.

[0016] In one embodiment, the amount of arginine or N-acetylcysteine ​​in the pharmaceutical combination or composition described herein ranges from about 25 mg to about 400 mg. Specific embodiments of the amount of arginine or N-acetylcysteine ​​include, but are not limited to, about 25 mg to about 400 mg, about 25 mg to about 390 mg, about 25 mg to about 380 mg, about 25 mg to about 370 mg, about 25 mg to about 360 mg, about 25 mg to about 350 mg, about 25 mg to about 340 mg, about 25 mg to about 330 mg, about 25 mg to about 320 mg, about 25 mg to about 310 mg, about 25 mg to about 300 mg, about 25 mg to about 290 mg, about 25 mg to about 280 mg, about 25 mg to about 270 mg, and about 2 5mg to about 260mg, about 25mg to about 250mg, about 25mg to about 240mg, about 25mg to about 230mg, about 25mg to about 220mg, about 25mg to about 210mg, about 25mg to about 200mg, about 25mg to about 190mg, about 25mg to about 180mg, Approximately 25mg to approximately 170mg, approximately 25mg to approximately 160mg, approximately 25mg to approximately 150mg, approximately 25mg to approximately 140mg, approximately 25mg to approximately 130mg, approximately 25mg to approximately 120mg, approximately 25mg to approximately 110mg, approximately 25mg to approximately 100mg, approximately 25mg to approximately 90mg, Approximately 25mg to approximately 80mg, approximately 25mg to approximately 70mg, approximately 25mg to approximately 60mg, approximately 25mg to approximately 50mg, approximately 25mg to approximately 40mg, approximately 25mg to approximately 30mg, approximately 30mg to approximately 400mg, approximately 30mg to approximately 390mg, approximately 30mg to approximately 380mg, approximately 30m g ~ approx. 370 mg, approx. 30 mg ~ approx. 360 mg, approx. 30 mg ~ approx. 350 mg, approx. 30 mg ~ approx. 340 mg, approx. 30 mg ~ approx. 330 mg, approx. mg~280mg, 30mg~270mg, 30mg~260mg, 30mg~300mg, 30mg~240mg, 30mg~230mg, 30mg~220mg, 30mg~210mg, 30mg~200mg, approx. 30mg to about 190mg, about 30mg to about 180mg, about 30mg to about 170mg, about 30mg to about 160mg, about 30mg to about 150mg, about 30mg to about 140mg, about 30mg to about 130mg, about 30mg to about 120mg, about 30mg to about 110mg,about 30 mg to about 100 mg, about 30 mg to about 90 mg, about 30 mg to about 80 mg, about 30 mg to about 70 mg, about 30 mg to about 60 mg, about 30 mg to about 50 mg, about 30 mg to about 40 mg, about 40 mg to about 400 mg, about 40 mg to about 390 mg, about 40 mg to about 380 mg, about 40 mg to about 370 mg, about 40 mg to about 360 mg, about 40 mg to about 350 mg, about 40 mg to about 340 mg, about 40 mg to about 330 mg, about 40 mg to about 320 mg, about 40 mg to about 310 mg, about 40 mg to about 300 mg, about 40 mg to about 290 mg, about 40 mg to about 2 80 mg, about 40 mg to about 270 mg, about 40 mg to about 260 mg, about 40 mg to about 400 mg, about 40 mg to about 240 mg, about 40 mg to about 230 mg, about 40 mg to about 220 mg, about 40 mg to about 210 mg, about 40 mg to about 200 mg, about 40 mg to about 190 mg, about 40 mg to about 180 mg, about 40 mg to about 170 mg, about 40 mg to about 160 mg, about 40 mg to about 150 mg, about 40 mg to about 140 mg, about 40 mg to about 130 mg, about 40 mg to about 120 mg, about 40 mg to about 110 mg, about 40 mg to about 100 mg, about 40 mg to about 90 mg , about 40 mg to about 80 mg, about 40 mg to about 70 mg, about 40 mg to about 60 mg, about 40 mg to about 50 mg, about 50 mg to about 400 mg, about 50 mg to about 390 mg, about 50 mg to about 380 mg, about 50 mg to about 370 mg, about 50 mg to about 360 mg, about 50 mg to about 350 mg, about 50 mg to about 340 mg, about 50 mg to about 330 mg, about 50 mg to about 320 mg, about 50 mg to about 310 mg, about 50 mg to about 300 mg, about 50 mg to about 290 mg, about 50 mg to about 280 mg, about 50 mg to about 270 mg, about 50 mg to about 260 mg, about 50 mg to about 500 mg, about 50 mg to about 240 mg, about 50 mg to about 230 mg, about 50 mg to about 220 mg, about 50 mg to about 210 mg, about 50 mg to about 200 mg, about 50 mg to about 190 mg, about 50 mg to about 180 mg, about 50 mg to about 170 mg, about 50 mg to about 160 mg, about 50 mg to about 150 mg, about 50 mg to about 140 mg, about 50 mg to about 130 mg, about 50 mg to about 120 mg, about 50 mg to about 110 mg, about 50 mg to about 100 mg, about 50 mg to about 90 mg, about 50 mg to about 80 mg, about 50 mg to about 70 mg, about 50 mg to about 60 mg,about 75 mg to about 400 mg, about 75 mg to about 390 mg, about 75 mg to about 380 mg, about 75 mg to about 370 mg, about 75 mg to about 360 mg, about 75 mg to about 350 mg, about 75 mg to about 340 mg, about 75 mg to about 330 mg, about 75 mg to about 320 mg, about 75 mg to about 310 mg, about 75 mg to about 300 mg, about 75 mg to about 290 mg, about 75 mg to about 280 mg, about 75 mg to about 270 mg, about 75 mg to about 260 mg, about 75 mg to about 750 mg, about 75 mg to about 240 mg, about 75 mg to about 230 mg, about 75 mg to about 220 mg, about 75 mg to about 210 mg, about 75 mg to about 200 mg, about 75 mg to about 190 mg, about 75 mg to about 180 mg, about 75 mg to about 170 mg, about 75 mg to about 160 mg, about 75 mg to about 150 mg, about 75 mg to about 140 mg, about 75 mg to about 130 mg, about 75 mg to about 120 mg, about 75 mg to about 110 mg, about 75 mg to about 100 mg, about 75 mg to about 90 mg, about 75 mg to about 80 mg, about 100 mg to about 400 mg, about 100 mg to about 390 mg, about 100 mg to about 380 mg, about 100 mg to about 370 mg, about 100 mg to about 360 mg, about 100 mg to about 350 mg, about 100 mg to about 340 mg, about 100 mg to about 330 mg, about 100 mg to about 320 mg, about 100 mg to about 310 mg, about 100 mg to about 300 mg, about 100 mg to about 290 mg, about 100 mg to about 280 mg, about 100 mg to about 270 mg, about 100 mg to about 260 mg, about 100 mg to about 1000 mg, about 100 mg to about 240 mg, about 100 mg to about 230 mg, about 100 mg to about 220 mg, about 100 mg to about 210 mg, about 100 mg to about 200 mg, about 100 mg to about 190 mg, about 100 mg g to about 180 mg, about 100 mg to about 170 mg, about 100 mg to about 160 mg, about 100 mg to about 150 mg, about 100 mg to about 140 mg, about 100 mg to about 130 mg, about 100 mg to about 120 mg, about 100 mg to about 110 mg, about 125 mg to about 400 mg, about 125 mg to about 390 mg, about 125 mg to about 380 mg, about 125 mg to about 370 mg, about 125 mg to about 360 mg, about 125 mg to about 350 mg, about 125 mg to about 340 mg, about 125 mg to about 330 mg, about 125 mg to about 320 mg, about 125 mg to about 310 mg,about 125 mg to about 300 mg, about 125 mg to about 290 mg, about 125 mg to about 280 mg, about 125 mg to about 270 mg, about 125 mg to about 260 mg, about 125 mg to about 1250 mg, about 125 mg to about 240 mg, about 125 mg to about 230 mg, about 125 mg to about 220 mg, about 125 mg to about 210 mg, about 125 mg to about 200 mg, about 125 mg to about 190 mg, about 125 mg to about 180 mg, about 125 mg to about 170 mg, about 125 mg to about 160 mg, about 125 mg to about 150 mg, about 125 mg to about 140 mg, about 125 mg to about 1 30mg, about 150mg to about 400mg, about 150mg to about 390mg, about 150mg to about 380mg, about 150mg to about 370mg, about 150mg to about 360mg, about 150mg to about 350mg, about 150mg to about 340mg, about 150mg to about 330mg, about 150mg to about 320mg, about 150mg to about 310mg, about 150mg to about 300mg, about 150mg to about 290mg, about 150mg to about 280mg, about 150mg to about 270mg, about 150mg to about 260mg, about 150mg to about 250mg, about 150mg to about 240mg, about 150mg to about 230mg, about 150mg to about 220mg, about 150mg to about 210mg, about 150mg to about 200mg, about 150mg to about 190mg, about 150mg to about 180mg, about 150mg to about 170mg, about 150mg to about 160mg, about 175mg to about 400mg, about 175mg to about 390mg, about 175mg to about 380mg, about 175mg to about 370mg, about 175mg to about 360mg, about 175mg to about 350mg, about 175mg to about 340mg, about 175mg to about 330mg, about 175mg to about 320mg, about 175mg to about 310mg, about 175mg to about 400mg. 5mg to about 300mg, about 175mg to about 290mg, about 175mg to about 280mg, about 175mg to about 270mg, about 175mg to about 260mg, about 175mg to about 250mg, about 175mg to about 240mg, about 175mg to about 230mg, about 175mg to about 220mg, about 175mg to about 210mg, about 175mg to about 200mg, about 175mg to about 190mg, about 175mg to about 180mg, about 200mg to about 400mg, about 200mg to about 390mg, about 200mg to about 380mg, about 200mg to about 370mg, about 200mg to about 360mg,about 200 mg to about 350 mg, about 200 mg to about 340 mg, about 200 mg to about 330 mg, about 200 mg to about 320 mg, about 200 mg to about 310 mg, about 200 mg to about 300 mg, about 200 mg to about 290 mg, about 200 mg to about 280 mg, about 200 mg to about 270 mg, about 200 mg to about 260 mg, about 200 mg to about 2000 mg, about 200 mg to about 240 mg, about 200 mg to about 230 mg, about 200 mg to about 220 mg, about 200 mg to about 210 mg, about 225 mg to about 400 mg, about 225 mg to about 390 mg, about 225 mg to about 3 80mg, about 225mg to about 370mg, about 225mg to about 360mg, about 225mg to about 350mg, about 225mg to about 340mg, about 225mg to about 330mg, about 225mg to about 320mg, about 225mg to about 310mg, about 225mg to about 300mg, about 225mg to about 290mg, about 225mg to about 280mg, about 225mg to about 270mg, about 225mg to about 260mg, about 225mg to about 250mg, about 225mg to about 240mg, about 225mg to about 230mg, about 225mg to about 220mg, about 225mg to about 210mg, about 225mg to about 200 mg, about 250 mg to about 400 mg, about 250 mg to about 390 mg, about 250 mg to about 380 mg, about 250 mg to about 370 mg, about 250 mg to about 360 mg, about 250 mg to about 350 mg, about 250 mg to about 340 mg, about 250 mg to about 330 mg, about 250 mg to about 320 mg, about 250 mg to about 310 mg, about 250 mg to about 300 mg, about 250 mg to about 290 mg, about 250 mg to about 280 mg, about 250 mg to about 270 mg, about 250 mg to about 260 mg, about 275 mg to about 400 mg, about 275 mg to about 390 mg, about 270 mg 5mg to about 380mg, about 275mg to about 370mg, about 275mg to about 360mg, about 275mg to about 350mg, about 275mg to about 340mg, about 275mg to about 330mg, about 275mg to about 320mg, about 275mg to about 310mg, about 275mg to about 300mg, about 275mg to about 290mg, about 275mg to about 280mg, about 300mg to about 400mg, about 300mg to about 390mg, about 300mg to about 380mg, about 300mg to about 370mg, about 300mg to about 360mg, about 300mg to about 350mg, about 300mg to about 340mg,Examples include about 300 mg to about 330 mg, about 300 mg to about 320 mg, about 300 mg to about 310 mg, about 325 mg to about 400 mg, about 325 mg to about 390 mg, about 325 mg to about 380 mg, about 325 mg to about 370 mg, about 325 mg to about 360 mg, about 350 mg to about 350 mg, about 325 mg to about 340 mg, about 325 mg to about 330 mg, about 350 mg to about 400 mg, about 350 mg to about 390 mg, about 350 mg to about 380 mg, about 350 mg to about 370 mg, about 350 mg to about 360 mg, about 375 mg to about 400 mg, about 375 mg to about 390 mg, and about 375 mg to about 380 mg.

[0017] In one embodiment, the pharmaceutical combination or composition described herein comprises about 0.5 mg to about 250 mg of a PDE5 inhibitor, about 25 mg to about 400 mg of arginine, and about 25 mg to about 400 mg of N-acetylcysteine. Certain embodiments of the pharmaceutical combination or composition described herein have the amounts of a PDE5 inhibitor, N-acetylcysteine, and arginine disclosed herein.

[0018] In some embodiments, the amount of PDE5 inhibitor in the pharmaceutical combination or composition described herein is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 110, 111, 112, 113, 114, 115, 116, 117, 118, 12 2, 43, 44, 45, 46, 47, 48, 49, or 50 mg, and the amount of arginine is about 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254 , 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, or 275 mg, and / or the amount of N-acetylcysteine ​​is about 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, or 275 mg. In certain embodiments, the amount of the PDE5 inhibitor is about 20 mg, the amount of arginine is about 250 mg, and / or the amount of N-acetylcysteine ​​is about 250 mg. In certain embodiments, the amount of the PDE5 inhibitor is about 20 mg, the amount of arginine is about 250 mg, and the amount of N-acetylcysteine ​​is about 250 mg.

[0019] In one embodiment, the daily dose of the pharmaceutical combination or composition is twice the dose of the PDE5 inhibitor, arginine, and / or N-acetylcysteine ​​described herein.In one embodiment, the pharmaceutical combination or composition disclosed herein is administered twice a day, so the daily dose of the pharmaceutical combination or composition is twice the dose of the PDE5 inhibitor, arginine, and / or N-acetylcysteine ​​described herein.

[0020] In some embodiments, the daily dose of the PDE5 inhibitor in the pharmaceutical combination or composition disclosed herein is about 4 mg to about 100 mg. In some embodiments, the daily dose of arginine in the pharmaceutical combination or composition disclosed herein is about 20 mg to about 1000 mg. In some embodiments, the daily dose of N-acetylcysteine ​​in the pharmaceutical combination or composition disclosed herein is about 20 mg to about 1000 mg. Certain embodiments of the pharmaceutical combination or composition disclosed herein have a daily dose of the PDE5 inhibitor of about 40 mg. Certain embodiments of the pharmaceutical combination or composition disclosed herein have a daily dose of arginine of about 500 mg. Certain embodiments of the pharmaceutical combination or composition disclosed herein have a daily dose of N-acetylcysteine ​​of about 500 mg. Certain embodiments of the pharmaceutical combinations or compositions disclosed herein have a daily dose of the PDE5 inhibitor of about 40 mg, a daily dose of arginine of about 500 mg, and / or a daily dose of N-acetylcysteine ​​of about 500 mg.

[0021] In one embodiment, the pharmaceutical combination or composition described herein is in a liquid formulation, wherein the combination of compounds disclosed herein has a sildenafil concentration of about 1-1000 nM, an arginine concentration of 1-1000 μM, and / or an N-acetylcysteine ​​concentration of 0.1-100 mM.

[0022] In one embodiment, the combinations or compositions disclosed herein are fast acting and / or can be taken over an extended period of time with fewer side effects.

[0023] In one embodiment, the combination or composition disclosed herein is in the form of a solid or liquid formulation, particularly a capsule or tablet or solution for injection or oral administration.

[0024] The present disclosure also provides a method for treating vascular disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition or combination disclosed herein. Alternatively, the present disclosure relates to a pharmaceutical composition or combination disclosed herein for use in treating vascular disease. Alternatively, the present invention discloses the use of a combination in the preparation of a medicament for treating vascular disease, the combination comprising sildenafil, arginine, and N-acetylcysteine.

[0025] In one embodiment, NAC and arginine can enhance the effect of a PDE5 inhibitor and simultaneously reduce the adverse effects of the PDE5 inhibitor.Therefore, the present disclosure also provides a method for reducing the adverse effects of a PDE5 inhibitor, comprising administering a combination or composition of NAC and arginine together with a PDE5 inhibitor.

[0026] In one embodiment, the dosages or administration amounts disclosed herein are effective or therapeutically effective amounts.

[0027] In one embodiment, the vascular disease of the present disclosure is selected from erectile dysfunction, altitude sickness, pulmonary hypertension, vascular fibrosis, and vascular sclerosis.

[0028] In one embodiment, the PDE5 inhibitor disclosed herein or a pharmaceutically acceptable salt thereof, arginine, and N-acetylcysteine ​​are administered simultaneously, sequentially, or separately.

[0029] In one embodiment, a PDE5 inhibitor disclosed herein, or a pharmaceutically acceptable salt thereof, is administered simultaneously with N-acetylcysteine.

[0030] In a further embodiment, the PDE5 inhibitor is sildenafil or sildenafil citrate.

[0031] In one embodiment, the combination or composition disclosed herein provides a synergistic effect compared to a PDE5 inhibitor or a pharmaceutically acceptable salt thereof, such as sildenafil or sildenafil citrate, alone.

[0032] In one embodiment, the combination or composition disclosed herein is for oral administration.

[0033] In one embodiment, the combination or composition disclosed herein is administered once, twice, or three times daily; or once, twice, three, four, five, six, or seven times weekly. In certain embodiments, the pharmaceutical combination or composition disclosed herein is administered twice daily.

[0034] In one embodiment, the agents (PDE5 inhibitor, arginine, and NAC) in the combinations or compositions disclosed herein are derivatives of said agents, such as, but not limited to, salts, pharmaceutically acceptable salts, solvates, hydrates, esters, tautomers, stereoisomers, enantiomers, or diastereomers thereof. Unless otherwise specified, the agents in the combinations or compositions disclosed herein include the aforementioned derivatives. [Brief explanation of the drawings]

[0035] [Figure 1] FIG. 1 is a scale chart of the relationship between the amount of NO released and treatment time in the treatment of vascular endothelial cells with sildenafil (SILD). [Figure 2] Figure 2 is a scale chart of NO release from vascular endothelial cells after treatment with sildenafil (SILD), sildenafil + N-acetylcysteine ​​(SN), and sildenafil + arginine + N-acetylcysteine ​​(SAN) for 10 minutes. [Figure 3]Figure 3 is a scale chart of the expression levels of endothelial nitric oxide synthase after treating endothelial cells with sildenafil (SILD), sildenafil + N-acetylcysteine ​​(SN), and sildenafil + arginine + N-acetylcysteine ​​(SAN) for 10 minutes. [Figure 4] FIG. 4 is a scale chart showing the relationship between the amount of cGMP expression in smooth muscle cells and treatment time in the treatment of smooth muscle cells with sildenafil (SILD). [Figure 5] FIG. 5 is a scale chart of cGMP expression in smooth muscle cells after treatment with sildenafil (SILD), sildenafil plus N-acetylcysteine ​​(SN), and sildenafil plus arginine plus N-acetylcysteine ​​(SAN) for 10 minutes. [Figure 6] FIG. 6 is a scale chart of endothelial cell free radical content after treatment of endothelial cells with sildenafil (SILD), sildenafil + N-acetylcysteine ​​(SN), and sildenafil + arginine + N-acetylcysteine ​​(SAN) for 10 minutes. [Figure 7] FIG. 7 is a scale chart of the vascular layer fibrosis index after treating vascular endothelial cells with sildenafil (SILD), sildenafil + N-acetylcysteine ​​(SN), and sildenafil + arginine + N-acetylcysteine ​​(SAN) for 10 minutes. [Figure 8] FIG. 8 is a scale chart of vascular permeability after treating vascular endothelial cells with sildenafil (SILD), sildenafil + N-acetylcysteine ​​(SN), and sildenafil + arginine + N-acetylcysteine ​​(SAN) for 10 minutes. DETAILED DESCRIPTION OF THE INVENTION

[0036] As used herein, "treatment" refers to a clinical intervention that seeks to alter the natural course of the individual or cell being treated and can be performed either prophylactically or during the course of clinical pathology. Desirable effects of treatment include prevention of disease onset or recurrence, alleviation of symptoms, reduction of any direct or indirect pathological consequences of disease, prevention or reduction of inflammation and / or tissue / organ damage, reduction in disease incidence, improvement or mitigation of the disease state, and remission or improved prognosis.

[0037] As used herein as adjectives, the term "pharmaceutical" or "pharmaceutically acceptable" means substantially non-toxic and substantially non-harmful to a recipient. A "pharmaceutical composition" further means that the carriers, solvents, excipients, and salts must be compatible with the active ingredients of the composition (e.g., the compounds of the present invention). Those skilled in the art will appreciate that the terms "pharmaceutical formulation" and "pharmaceutical composition" are generally interchangeable and will be used as such for purposes of this application.

[0038] An "individual" or "subject" as disclosed herein is a vertebrate. In certain embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, farm animals (e.g., cows), sport animals, pets (e.g., cats, dogs, and horses), primates, mice, and rats. In certain embodiments, the vertebrate is a human.

[0039] As disclosed herein, the term "effective amount" refers to an amount effective to achieve a desired therapeutic or preventive result at a specific dosage and for a period of time necessary. In one embodiment, the PDE5 inhibitor, arginine, and / or NAC in the combination or composition disclosed herein are in an effective amount.

[0040] The "therapeutically effective amount" of a substance / molecule disclosed herein may vary depending on factors such as the disease state, age, sex, and weight of the individual, as well as the ability of the substance / molecule to induce a desired response in the individual. A therapeutically effective amount is also an amount in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the substance / molecule. A "prophylactically effective amount" refers to an amount effective to achieve the desired preventive result at a certain dosage and for a necessary period of time. Typically, but not necessarily, the prophylactically effective amount will be less than the therapeutically effective amount, since a prophylactic dose is used in subjects before or at an earlier stage of disease. In one embodiment, the PDE5 inhibitor, arginine, and / or NAC in the combination or composition disclosed herein are in a therapeutically effective amount.

[0041] As disclosed herein, administration "in combination with" includes simultaneous (concurrent) administration and sequential administration in any order. As used herein, "combination therapy" or "in combination" includes administration of a PDE5 inhibitor, arginine, and NAC as part of a specific therapeutic regimen intended to provide beneficial effects from the synergistic action of these agents.

[0042] "Excipient," as used herein, includes a pharmaceutically acceptable carrier or stabilizer that is nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. In one embodiment, the physiologically acceptable carrier is an aqueous pH buffered solution.

[0043] The term "about" as used herein refers to a variation of about ±1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%.

[0044] The present disclosure has surprisingly found that the combination or composition of a PDE5 inhibitor (preferably sildenafil), arginine and N-acetylcysteine ​​can increase the reaction rate of the PDE5 inhibitor or reduce the reaction time of the PDE5 inhibitor, and / or significantly increase the expression of intracellular nitric oxide synthase, compared with the administration of the PDE5 inhibitor alone.Therefore, the present disclosure provides a pharmaceutical combination comprising a PDE5 inhibitor (preferably sildenafil), arginine and N-acetylcysteine, and the method of using the combination or composition to effectively treat vascular disease with fewer side effects.

[0045] Several phosphodiesterase type 5 (PDE5) inhibitors or their pharmaceutically acceptable salts, such as sildenafil (SLID), tadalafil, and vardenafil, have been approved by health authorities for the treatment of erectile dysfunction. Previous experiments have shown that the mechanism by which penile erection occurs is that nitric oxide (NO) diffuses from vascular endothelial cells into penile smooth muscle cells, activating guanylate cyclase (GC), thereby increasing the production of cyclic guanosine monophosphate (cGMP) and inhibiting calcium ion influx, which reduces the intracellular calcium ion concentration and relaxes the smooth muscle of the corpus cavernosum. The corpus cavernosum then becomes congested with a large amount of blood, resulting in erection. For example, sildenafil inhibits the hydrolysis of cGMP, allowing cGMP to continuously act to achieve the effects of congestive heart failure and erection. According to the package insert for sildenafil, sildenafil does not have a direct relaxant effect on isolated human corpus cavernosum, but enhances the effects of nitric oxide (NO) by inhibiting phosphodiesterase type 5 (PDE5), which is responsible for the breakdown of cGMP in the corpus cavernosum. However, sildenafil also has several side effects. After administration, sildenafil can cause headache, abdominal pain, diarrhea, flushing, dizziness, weakness, pruritus, and erythema. Most side effects occur within minutes of taking sildenafil. Long-term use of sildenafil can lead to severe hypertension, which can increase problems such as temporary loss of vision and coronary artery sclerosis. These side effects are generally thought to be caused by vasodilation; rapid vasodilation can also expose vascular endothelial cells to pressure from high blood flow, causing them to release endogenous free radicals, which in turn damage vascular cells.

[0046] Arginine is a drug that can be used as oral tablets and oral capsules.In the erectile process, L-arginine can catalyze the production of nitric oxide by endothelial nitric oxide synthase, and nitric oxide can be rapidly released into the space between cavernous tissues, thereby reducing the amount of divalent calcium ions in cavernous artery smooth muscle cells, causing smooth muscle relaxation and increasing blood flow.

[0047] N-acetylcysteine ​​(NAC) has antioxidant properties and can promote the intracellular conversion of glutathione disulfide (GSSG) to glutathione (GSH), which is an essential component for the synthesis of the body's most effective antioxidant enzymes. Furthermore, NAC has been reported to have an inhibitory effect on inducible nitric oxide synthase (NOS) protein and NO production (Araki et al., N-acetylcysteine ​​inhibits induction of nitric oxide synthase in 3T3-L1 adipocytes, J UOEH 29 (4):417-429 (2007); Bergamini et al., N-acetylcysteine ​​inhibits in vivo nitric oxide production by inducible nitric oxide synthase, NITRIC OXIDE: Biology and Chemistry, Vol. 5, No. 4, pp. 349-360 (2001); Rota et al., N-acetylcysteine ​​negatively modulates nitric oxide production in endotoxin-treated rats through inhibition of NF-κB activation; Antioxidants & Redox Signaling, Vol. 4, No. 1 (2004)). Thus, NAC causes a detrimental effect on NO production, opposite to the effect produced by arginine.

[0048] However, the present disclosure has unexpectedly found that a combination of NAC and arginine can enhance the effects of PDE5 inhibitors while simultaneously reducing their adverse effects.

[0049] In the pharmaceutical combination, examples of the amount or dose of the PDE5 inhibitor used herein include, but are not limited to, about 1 mg to about 500 mg, about 5 mg to about 490 mg, about 5 mg to about 450 mg, about 5 mg to about 400 mg, about 5 mg to about 350 mg, about 5 mg to about 300 mg, about 5 mg to about 250 mg, about 5 mg to about 200 mg, about 5 mg to about 175 mg, about 5 mg to about 150 mg, about 5 mg to about 125 mg, about 5 mg to about 100 mg, about 5 mg to about 90 mg, about 5 mg to about 80 mg, about 5 mg to about 70 mg, about 5 mg to about 60 mg, about 5 mg to about 50 mg, About 5 mg to about 40 mg, about 5 mg to about 30 mg, about 5 mg to about 20 mg, about 5 mg to about 10 mg, about 10 mg to about 480 mg, about 10 mg to about 470 mg, about 10 mg to about 460 mg, about 10 mg to about 450 mg, about 10 mg to about 440 mg, about 10 mg to about 430 mg, about 10 mg~420mg, 10mg~410mg, 10mg~400mg, 10mg~390mg, 10mg~380mg, 10mg~370mg, 10mg~360mg, 10mg~350mg, 10mg~340mg, 10mg~330m g, approx. 10 mg ~ approx. 320 mg, approx. 10 mg ~ approx. 310 mg, approx. 10 mg ~ approx. 300 mg, approx. 10 mg ~ approx. 290 mg, approx. 10 mg ~ approx. 280 mg, approx. Approx. 230 mg, approx. 10 mg ~ approx. 220 mg, approx. 10 mg ~ approx. 210 mg, approx. 10 mg ~ approx. 200 mg, approx. 10 mg ~ approx. 190 mg, approx. 10mg to about 130mg, about 10mg to about 120mg, about 10mg to about 110mg, about 10mg to about 100mg, about 10mg to about 90mg, about 10mg to about 80mg, about 10mg to about 70mg, about 10mg to about 60mg, about 10mg to about 50mg, about 10mg to about 40mg, about 1 0mg to about 30mg, about 10mg to about 20mg, about 20mg to about 450mg, about 20mg to about 400mg, about 20mg to about 350mg, about 20mg to about 300mg, about 20mg to about 250mg, about 20mg to about 200mg, about 20mg to about 175mg, about 20mg to about 150mg,About 20mg to about 125mg, about 20mg to about 100mg, about 20mg to about 90mg, about 20mg to about 80mg, about 20mg to about 70mg, about 20mg to about 60mg, about 20mg to about 50mg, about 20mg to about 40mg, about 20mg to about 30mg, about 50mg to about 450mg, about 50 mg ~ about 400mg, about 50mg - about 350mg, about 50mg - about 300mg, about 50mg - about 250mg, about 50mg - about 200mg, about 50mg - about 175mg, about 50mg - about 150mg, about 50mg - about 125mg, about 50mg - about 100mg, about 50mg - about 90mg, Examples include about 50 mg to about 80 mg, about 50 mg to about 70 mg, about 50 mg to about 60 mg, about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 100 mg to about 175 mg, about 100 mg to about 150 mg, about 100 mg to about 125 mg, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg.

[0050] In the pharmaceutical combination, examples of the amount or dose of arginine or N-acetylcysteine ​​used herein include, but are not limited to, about 10 mg to about 2 g, about 10 mg to about 1.9 g, about 10 mg to about 1.8 g, about 10 mg to about 1.7 g, about 10 mg to about 1.6 g, about 10 mg to about 1.5 g, about 10 mg to about 1.4 g, about 10 mg to about 1.3 g, about 10 mg to about 1.2 g, about 10 mg to about 1.1 g, about 10 mg to about 1.2 g, about 10 mg to about 1.3 g, about 10 mg to about 1.4 g, about 10 mg to about 1.5 g, about 10 mg to about 1.6 g, about 10 mg to about 1.7 g, about 10 mg to about 1.8 g, about 10 mg to about 1.9 g, about 10 mg to about 1.9 g, about 10 mg to about 1.8 ...1 g, about 10 mg to about 1.2 g, about 10 mg to about 1.3 g, about 10 mg to about 1.4 g, about 10 mg to about 1.5 g, about 10 mg to about 1.6 g, about 10 mg to about 1.7 g, about 10 mg to about 1.8 g, about 10 mg to about 1.9 g mg ~ about 1.0g, about 10mg - about 900mg, about 10mg - about 800mg, about 10mg - about 700mg, about 10mg - about 600mg, about 10mg - about 500mg, about 10mg - about 400mg, about 10m g ~ about 300mg, about 10mg - about 200mg, about 10mg - about 100mg, about 10mg - about 50mg, about 10mg - about 25mg, about 100mg - about 2g, about 100mg - about 1.9g, about 100mg - about 1 .8g, about 100mg to about 1.7g, about 100mg to about 1.6g, about 100mg to about 1.5g, about 100mg to about 1.4g, about 100mg to about 1.3g, about 100mg to about 1.2g, about 100mg to about 1 .1g, about 100mg to about 1.0g, about 100mg to about 900mg, about 100mg to about 800mg, about 100mg to about 700mg, about 100mg to about 600mg, about 100mg to about 500mg, about 100m g to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 200 mg, or about 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, or 1.0 g, or about 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, 100, 50, 25, or 10 mg.

[0051] The combinations or compositions disclosed herein generally contain one or more pharmaceutically acceptable excipients, carriers, or diluents. The specific carriers, diluents, or excipients used will depend on the means and purpose for which the active ingredient is being administered. Tablet formulations generally contain substances such as diluents, binders, lubricants, disintegrants, and mixtures thereof. Suitable diluents include various types of starch, lactose, mannitol, kaolin, calcium phosphate or calcium sulfate, inorganic salts (e.g., sodium chloride), powdered sugar, and powdered cellulose derivatives. More specifically, examples of diluents or fillers include lactose, mannitol, xylitol, dextrose, sucrose, sorbitol, compressible sugar, microcrystalline cellulose, powdered cellulose, starch, pregelatinized starch, dextrates, dextran, dextrin, dextrose, maltodextrin, calcium carbonate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, magnesium carbonate, magnesium oxide, poloxamers such as polyethylene oxide, and hydroxypropyl methylcellulose. If desired, a binder may be added. Suitable binders include cellulose (e.g., cellulose, methylcellulose, ethylcellulose, and hydroxymethylcellulose), polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, starch, sugars (e.g., lactose, sucrose, fructose, and glucose), natural and synthetic gums (e.g., acacia, alginate, and gum arabic), and waxes. Lubricants are typically used in tablet formulations to prevent the tablet and punches from sticking in the die. Suitable lubricants include calcium stearate, glyceryl monostearate, glyceryl palmitostearate, hydrogenated vegetable oil, light mineral oil, magnesium stearate, mineral oil, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc, and zinc stearate. A disintegrant may be added to the combination or composition to break up the dosage form and release the compound.Suitable disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, powdered cellulose, lower alkyl-substituted hydroxypropyl cellulose, polacrilin potassium, starch, pregelatinized starch, and sodium alginate.

[0052] The combination or composition of a PDE5 inhibitor combined with arginine and NAC is useful for treating vascular diseases.Surprisingly, the combination or composition of NAC and arginine can enhance the effect of PDE5 inhibitors and simultaneously reduce the adverse effects of PDE5 inhibitors.Therefore, the present disclosure provides a method for treating vascular diseases, comprising administering to a subject an effective amount of a pharmaceutical combination or composition of a PDE5 inhibitor, arginine and NAC.The present disclosure also provides a method for treating diseases associated with PDE5 inhibition while reducing the adverse effects of PDE5 inhibition, comprising administering to a subject an effective amount of NAC and arginine in combination with a PDE5 inhibitor.

[0053] Examples of vascular diseases include, but are not limited to, erectile dysfunction, altitude sickness, pulmonary hypertension, vascular fibrosis, and vascular sclerosis.

[0054] The beneficial effect of the combination includes, but is not limited to, pharmacokinetic and / or pharmacodynamic synergy resulting from the combination of the agents. Administration of the agents in the combination typically occurs over a predetermined period of time (usually minutes, hours, days, or weeks, depending on the combination selected). Combination therapy is intended to encompass not only sequential administration of the indicated therapeutic agents, i.e., administration of each therapeutic agent at different times, but also substantially simultaneous administration of these therapeutic agents. Administration can be achieved, for example, by administering to the subject a single oral dosage form having a fixed ratio of each agent, or multiple, single oral dosage forms for each of the therapeutic agents. Sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any appropriate route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissue.

[0055] In another embodiment, the combinations and compositions of the present disclosure can be administered parenterally (e.g., intramuscularly, intrathecally, intravenously, and intraarterially), preferably intravenously. Typically, the compounds and compositions of the present disclosure for intravenous administration are solutions in sterile isotonic aqueous vehicles, such as water, saline, Ringer's solution, or dextrose solution. If necessary, the compositions may also contain a solubilizing agent. Combinations and compositions for intravenous administration may optionally contain a local anesthetic, such as lignocaine, to ease pain at the injection site. For intravenous administration, the combinations and compositions of the present disclosure can be supplied as a sterile, dry, lyophilized powder or water-free concentrate in a hermetically sealed container, such as an ampoule or sachet, indicating the quantity of active agent. Such powders or concentrates are then diluted with an appropriate aqueous medium prior to intravenous administration. The powders or concentrates can be provided in ampoules of sterile water, saline solution, or other appropriate aqueous medium for dilution prior to administration. Alternatively, the combinations and compositions of the present disclosure can be supplied in a ready-to-administer, premixed form. When the combination and composition of the present disclosure are administered by intravenous infusion, they can be dispensed, for example, with an infusion bottle containing sterile pharmaceutical grade water, saline solution, or another suitable medium.Rectal administration can be achieved by using suppositories formulated from conventional carriers such as cocoa butter, modified vegetable oils, and other fat bases.Suppositories can be formulated by well-known methods using well-known formulations; for example, see Remington: The Science and Practice of Pharmacy, edited by Alfonso R. Gennaro, Mack Publishing Co. Easton, Pa., 19th edition, 1995, pages 1591-1597, which is incorporated herein by reference.

[0056] Therapeutically effective dosage regimens for treating a particular disorder or condition will depend on its nature and severity and can be determined by standard clinical techniques according to the judgment of the physician. Moreover, in vitro or in vivo assays can be used to help identify optimal dosages. Of course, the amount of a compound of the present invention that constitutes a therapeutically effective dose will also depend on the route of administration. [Example]

[0057] The following examples are provided by way of illustration and are not intended to limit the scope of the present invention.

[0058] Example Experimental Method Drug preparation Sildenafil, arginine, N-acetylcysteine, and S-nitroso-N-acetylpenicillamine (SNAP) are dissolved and diluted in phosphate buffered saline (PBS). In particular, the following solutions are prepared: a solution containing 100 nM sildenafil (hereinafter referred to as SILD or "S"); a mixed solution of 100 nM sildenafil and 10 mM N-acetylcysteine ​​(hereinafter referred to as "SN"); a mixed solution of 100 nM sildenafil, 150 μM arginine, and 10 mM N-acetylcysteine ​​(hereinafter referred to as "SAN"); a mixed solution of 100 nM sildenafil and 100 μM S-nitroso-N-acetylpenicillamine (SNAP) (hereinafter referred to as "SS"); and a mixed solution of 100 nM sildenafil, 100 μM SNAP, and 10 mM N-acetylcysteine ​​(hereinafter referred to as "SSN"). The solutions are stored at a temperature of 4°C. For clinical trials and human studies, a SAN combination containing 20 mg sildenafil, 250 mg arginine, and 250 mg NAC will be used and administered to subjects.

[0059] cell culture Human umbilical vein endothelial cells (HUVECs) obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan) were cultured in Ham's F-12K (Sigma-Aldrich) medium containing endothelial cell growth supplement (Millipore), heparin (Sigma-Aldrich), 2.2 mg / mL sodium bicarbonate, and 10% FBS (Gibco). HUVECs at passages 5–15 were used for all applications. Human pulmonary artery vascular smooth muscle (PAVSM) cells and growth medium were obtained from Lonza Inc. (Walkersville, MD, USA). Cells were grown at 37°C in a 5% CO2 atmosphere.

[0060] Cell treatment For cell treatment, HUVEC and PAVSM cells were grown to confluence in the upper and lower wells of a Transwell® unit, respectively. The culture medium volume in each upper chamber was 1.5 mL, and the culture medium volume in each lower chamber was 2.5 mL. Confluent HUVEC cells were treated with normal medium in the presence or absence of S, SS, SN, SSN, and SAN for 0, 10, 20, 30, and 60 minutes. All HUVEC and PAVSM supernatants were collected immediately after treatment for NO analysis. HUVEC and PAVSM cells were rinsed in ice-cold phosphate-buffered saline and then solubilized in ice-cold 6% trichloroacetic acid for protein extraction. Samples were stored at -80°C until assay.

[0061] Analysis of nitric oxide release from endothelial cells Nitric oxide levels were assessed using an NO assay kit (Promega). After treatment, the supernatants of HUVECs were collected for the NO assay reagent kit, and 200 μL was applied per well according to the manufacturer's instructions. The relative luminance units (RLU) emitted by the product were measured using a microreader.

[0062] Analysis of endothelial nitric oxide synthase expression levels Endothelial nitric oxide synthase (eNOS) activity was assessed using an eNOS assay kit (Promega). After treatment, the supernatant plates were removed from the incubator and allowed to equilibrate to room temperature for approximately 30 minutes, after which the eNOS assay reagent (200 μL / well) was applied according to the manufacturer's instructions. The relative luminance units (RLU) emitted by the product were measured using a microreader.

[0063] Analysis of cGMP in smooth muscle cells Samples were treated with drugs for 0, 10, 20, 30, and 60 minutes, and cGMP and protein were measured as previously described (Garmaroudi et al., Systems Pharmacology and Rational Polypharmacy: Nitric Oxide-Cyclic GMP Signaling Pathway as an Illustrative Example and Derivation of the General Case; PLOS COMPUTATIONAL BIOLOGY, DOI: 10.1371 / journal.pcbi.1004822, March 17, 2016). cGMP formation was measured by immunoassay according to the cGMP assay (Cayman Chemical Co., Ann Arbor, MI).

[0064] Quantitative free radical ROS content detection method ROS detection studies were performed using the Cm-H2DCFDA ROS detection kit (Invitrogen, Grand Island, NY, USA). For cell-free ROS detection, PM at each concentration was used. 2.5 Aliquots of 100 μL serum-free medium containing α-H2DCFDA were pipetted into black 96-well plates and mixed with 10 μL Hank's Balanced Salt Solution (HBSS) containing Cm-H2DCFDA (final concentration 25 μM) activated by pre-incubation at 37°C for 30 min. ROS generation was measured every 10 min for up to 2 h using a microreader at 490 nm excitation and 530 nm emission. Values ​​were presented as RFU. For intracellular ROS detection, HUVECs were incubated in PM2.5 After incubation with Cm-H2DCFDA for 24 hours, they were washed with PBS and treated with fresh serum-free medium containing Cm-H2DCFDA (final concentration 25 μM). The samples were then immediately measured using a Promega microreader. 4 The ROS levels generated by surviving treated cells were expressed as a percentage of the ROS generated by the same number of surviving negative control cells.

[0065] Vascular fibrosis index assay: detection of free radical-induced lipid peroxidation (malondialdehyde, MDA) Lipid peroxidation in HUVEC protein extracts was quantified by measuring the concentration of MDA by spectrofluorometric assay using an "MDA assay kit" as described by Richard et al., 1992. Quantification was achieved by parallel measurement of a standard curve of known MDA concentrations, and results were expressed as a change in the level of control.

[0066] Vascular permeability analysis: VEGF-A release amount The amount of VEGFA was evaluated using a VEGFA assay kit (Promega). After treatment, the plate was removed from the incubator and allowed to equilibrate to room temperature for approximately 30 minutes, and then VEGFA assay reagent (200 μL / well) was applied according to the manufacturer's instructions. The relative absorbance units emitted by the product at 540 nm were measured using a microreader.

[0067] Studies of the effectiveness of SAN in humans A human / clinical study will be conducted in which patients with erectile dysfunction will be treated with a SAN combination containing 20 mg sildenafil, 250 mg arginine, and 250 mg NAC. The pharmacokinetics and pharmacodynamics of SAN, as well as the efficacy of SAN in treating ED and vascular disease, will be assayed. Evaluation indicators for ED and vascular disease known in the art will be applied herein, including assays for NO, NOS, ROS, cGMP, and indicators for vascular disease described herein or known in the art. Subjects receiving the SAN combination will be between 19 and 87 years of age. The therapeutic effect of SAN on ED will be assessed by a global assessment questionnaire, daily erection records, the International Institute of Erectile Function (IIEF, a validated sexual function questionnaire), and / or a sexual partner questionnaire.

[0068] Experimental results Example 1. Combination SAN increases the reaction rate of sildenafil and decreases the reaction time of sildenafil FIG. 1 is a scale chart showing the relationship between the amount of NO released by vascular endothelial cells treated with a drug or drug combination disclosed herein and treatment time. As shown in FIG. 1, the amount of nitric oxide released increases with SILD treatment time. Compared with the control group (0 minutes of drug treatment), the amount of nitric oxide released reaches approximately 1.6 times after 10 minutes of drug treatment, approximately 2.6 times after 20 minutes of drug treatment, approximately 3.1 times after 30 minutes of treatment, and approximately 3.5 times after 60 minutes of treatment. FIG. 2 is a scale chart showing the amount of nitric oxide released by vascular endothelial cells after 10 minutes of treatment with SILD, SN, and SAN. As shown in FIG. 2, after 10 minutes of drug treatment, the amount of nitric oxide released by the SILD group (S) reaches approximately 1.6 times, the amount of nitric oxide released by the SN group reaches approximately 2 times, and the amount of nitric oxide released by the SAN group reaches approximately 3.1 times, compared with the control group (no drug; considered as the reference). The results show that only 10 minutes of treatment with SAN can achieve the same NO release effect as 30 minutes of treatment with SILD; see Figures 1 and 2. That is, the reaction rate of SILD in the combination SAN is faster than that of SILD alone.

[0069] Example 2. Combination SAN increases nitric oxide synthase expression Figure 3 shows a scale chart of the expression levels of endothelial nitric oxide synthase (ENS) after 10 minutes of treatment of endothelial cells with SILD, SN, and SAN; SS and SSN were also used for treatment (results not shown in Figure 3). Compared with the control group (no drug; considered as reference), the expression levels of ENS in the SILD(S), SS, SN, and SAN groups were approximately 2-fold, 2.5-fold, 2.4-fold, and 2.8-fold higher, respectively. The results show that the expression levels of ENS in SAN treatment were significantly higher than those in SILD treatment alone; see Figure 3.

[0070] Example 3. Combination SAN increases the efficacy and response rate of sildenafil and decreases the response time of sildenafil Figure 4 is a scale chart showing the relationship between the cGMP expression level of smooth muscle cells and treatment time in the treatment of smooth muscle cells with sildenafil (SILD). The cGMP content of smooth muscle cells reflects the degree of vasodilation; as shown in Figure 4, the amount of cGMP increases with the duration of SILD treatment. Compared with the control group (0 minutes of drug treatment; considered as the reference), the amount of cGMP reaches approximately 2.6 times, 3 times, 4.7 times, and 5.5 times after 10, 20, 30, and 60 minutes of drug treatment, respectively.

[0071] Figure 5 shows a scale chart of the cGMP expression levels in smooth muscle cells after 10 minutes of treatment with SILD, SN, and SAN. As shown in Figure 5, after 10 minutes of treatment, the cGMP levels increased by approximately 2.6 times in the SILD (S) group, approximately 2.7 times in the SN group, and approximately 4.6 times in the SAN group compared to the control group (no drug; considered as the reference). The results show that the cGMP levels in smooth muscle cells increased after treating cells with SILD; when cells were treated with SAN, only 10 minutes of treatment achieved the same effect as 30 minutes of treatment with SILD (see Figures 4 and 5). That is, the drug response rate in SAN treatment was faster than that in SILD treatment alone.

[0072] Example 4. Combination SAN reduces vascular layer fibrosis index and vascular permeability induced by sildenafil Figure 6 is a scale chart showing the free radical content of vascular endothelial cells after 10 minutes of treatment with SILD, SN, and SAN. After treatment with SILD, vascular endothelial cells are subjected to pressure from high blood flow due to rapid vasodilation caused by SILD, which promotes the release of endogenous free radicals. Figure 6 shows that the amount of intracellular ROS changes with treatment with different drugs. Compared with the control group (no drug treatment; considered as reference), the amount of ROS is approximately 2.8-fold in the SILD(S) group and approximately 1-fold in the SN and SAN groups. The results indicate that the amount of ROS generated during treatment with SILD can be effectively reduced by the combination disclosed herein, particularly NAC in the combination.

[0073] Figure 7 is a scale chart showing the vascular layer fibrosis index after vascular endothelial cells were treated with SILD, SN, and SAN for 10 minutes. Increased endogenous free radicals cause peroxidation of cellular lipids, further leading to vascular fibrosis lesions. Figure 7 shows that the amount of intracellular malondialdehyde (MDA) changes after treatment with different drugs. Compared with the control group (no drug treatment; considered as reference), the amount of MDA is approximately 2.8 times higher in the SILD group and approximately 1 times higher in the SN and SAN groups. The results show that the amount of MDA produced by SILD is effectively reduced by the combination disclosed herein, particularly by NAC in the combination.

[0074] Figure 8 is a scale chart showing vascular permeability after 10 minutes of treatment of vascular endothelial cells with SILD, SN, and SAN. A local increase in vascular permeability is one of the indicators of vascular fibrosis and vascular sclerosis, and an increase in the vascular permeability factor VEGFA (vascular endothelial growth factor A) causes vascular fibrosis and vascular sclerosis. Figure 8 shows that the amount of intracellular VEGFA changes with treatment with different drugs. Compared with the control group (no drug treatment; considered as the reference), the amount of VEGFA reaches approximately 3.9 times in the SILD group (S) and approximately 1 time in the SN and SAN groups. The results show that the amount of VEGFA caused by treatment with SILD is effectively reduced by the combination disclosed herein, particularly NAC in the combination.

[0075] Example 5. A human / clinical study will be conducted in which patients with erectile dysfunction will be treated with a SAN combination containing 20 mg sildenafil, 250 mg arginine, and 250 mg NAC. The time at which peak blood concentrations can be observed when subjects orally ingest the SAN combination in a fasted state will be determined; previous data indicate that sildenafil alone is rapidly absorbed after administration (peak blood concentrations can be observed within 30 to 120 minutes (median 60 minutes)), with mean absolute bioavailability from oral administration ranging between 25% and 63%. The clearance rate of sildenafil will be observed, particularly in healthy elderly subjects (ages 65 and older); previous data have shown that after administration of sildenafil alone, the clearance rate in elderly subjects is reduced, with free plasma concentrations 40% higher than in younger subjects (ages 18 to 45). The efficacy and safety of SAN will be further evaluated in a randomized, double-blind, placebo-controlled trial. The subjects receiving the SAN combination are between 19 and 87 years old, and suffer from erectile dysfunction of various etiologies (organic, psychogenic, or mixed). The therapeutic effects of SAN are then evaluated using a global assessment questionnaire, daily erection records, the International Institute of Erectile Function (IIEF, a validated sexual function questionnaire), and a sexual partner questionnaire. Previous data indicate that the improvement rates after sildenafil monotherapy are 62% (25 mg), 74% (50 mg), and 82% (100 mg), respectively, compared with only 25% for placebo. Based on relevant preliminary results, the inventors believe that the SAN combination can achieve the same efficacy as known drugs with higher doses, faster response rates, and / or fewer side effects.

[0076] The results show that the SAN combination disclosed herein can significantly reduce the amount of ROS generated after treating smooth muscle cells with SILD, reduce ROS-induced cellular lipid peroxidation and VEGFA, and further reduce the abnormally increased vascular permeability and the risk of vascular sclerosis.

[0077] Furthermore, the SAN combination disclosed herein surprisingly increases the reaction rate of SILD, reduces the reaction time, and removes excess free radicals in cells, so as to achieve the function of protecting blood vessels and preventing fibrosis and hardening of blood vessel walls after long-term use. Therefore, the combination or composition disclosed herein for treating vascular diseases can be taken for a long period of time with fewer side effects.

[0078] The above description and examples only represent the description of the preferred embodiments of the present invention. Those skilled in the art can make modifications according to their general knowledge and the present invention herein, but these modifications should still be within the scope of the present invention herein in terms of the spirit of the present invention.

Claims

1. A pharmaceutical composition comprising a phosphodiesterase type 5 (PDE5) inhibitor or a pharmaceutically acceptable salt thereof, arginine, and N-acetylcysteine, and optionally a pharmaceutically acceptable excipient.

2. 2. The pharmaceutical composition of claim 1, wherein the PDE5 inhibitor is selected from sildenafil, tadalafil, and vardenafil.

3. the amounts of PDE5 inhibitor, arginine, and N-acetylcysteine ​​range from about 0.1% (w / w) to about 50% (w / w), from about 4.0% (w / w) to about 80% (w / w), and from about 4% (w / w) to about 80% (w / w), respectively; or 3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises from about 0.5 mg to about 250 mg of a PDE5 inhibitor, from about 25 mg to about 400 mg of arginine, and from about 25 mg to about 400 mg of N-acetylcysteine.

4. 3. The pharmaceutical composition of claim 1, further comprising one or more preservatives selected from the group consisting of antibacterial agents, antifungal agents, and disinfectants.

5. 3. The pharmaceutical composition according to claim 1 or 2, in the form of a solid or liquid formulation.

6. 6. The pharmaceutical composition of claim 5, which is a capsule or tablet.

7. 3. The pharmaceutical composition of claim 1 or 2 for use in treating a vascular disease in a subject in need thereof.

8. 8. The pharmaceutical composition for use according to claim 7, wherein the vascular disease is selected from erectile dysfunction, altitude sickness, pulmonary hypertension, vascular fibrosis, and vascular sclerosis.

9. The pharmaceutical composition of claim 7, wherein the composition increases the reaction rate of sildenafil or decreases the reaction time of sildenafil compared to administration of sildenafil alone.

10. The pharmaceutical composition of claim 7, wherein the composition significantly increases the expression of nitric oxide synthase in cells.

11. 8. The pharmaceutical composition of claim 7, wherein the PDE5 inhibitor or a pharmaceutically acceptable salt thereof, arginine, and N-acetylcysteine ​​are administered simultaneously, sequentially, or separately.

12. 8. The pharmaceutical composition according to claim 7, wherein the PDE5 inhibitor is sildenafil or sildenafil citrate.

13. The pharmaceutical composition of claim 7, wherein the composition is administered twice daily.

14. A pharmaceutical composition comprising an effective amount of NAC and arginine in combination with a PDE5 inhibitor or a pharmaceutically acceptable salt thereof for use in treating a disease associated with PDE5 inhibition while reducing the adverse effects of PDE5 inhibition.

15. The pharmaceutical composition described in claim 14, wherein the disease associated with the inhibition of PDE5 is a vascular disease; and the vascular disease is selected from erectile dysfunction, altitude sickness, pulmonary hypertension, vascular fibrosis, and vascular sclerosis.

16. The pharmaceutical composition of claim 14, wherein the combination increases the rate of response of sildenafil or decreases the time of response of sildenafil compared to administration of sildenafil alone.

17. The pharmaceutical composition of claim 14, wherein the composition significantly increases the expression of nitric oxide synthase in cells.

18. The pharmaceutical composition of claim 14, wherein the PDE5 inhibitor or a pharmaceutically acceptable salt thereof, arginine, and N-acetylcysteine ​​are administered simultaneously, sequentially, or separately.

19. The pharmaceutical composition described in claim 14, wherein the PDE5 inhibitor is sildenafil or sildenafil citrate.

20. The pharmaceutical composition of claim 14, wherein the composition is administered twice daily.

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