KR102116712B1 - Formulations for canine heart disease containing oral dosing formulations with highly palatable excipients - Google Patents

Abstract

The oral administration preparations with improved palatability of the present invention have improved palatability in canine animals, which facilitates long-term oral administration, and has the activity of treating or alleviating symptoms of heart disease, including bipolar valve degeneration, and improving blood circulation. .

Description

FORMULATIONS FOR CANINE HEART DISEASE CONTAINING ORAL DOSING FORMULATIONS WITH HIGHLY PALATABLE EXCIPIENTS}

The present invention relates to a preparation for treating or treating symptoms of bicuspid valve degeneration in a canine animal, including an oral administration preparation with improved palatability, and has an effect equal to or higher than that of a conventional product, while improving palatability, thereby facilitating oral administration. It relates to a pharmaceutical composition in powder form, a feed additive composition, and the like.

In the case of congestion, unlike a treatment for humans, no matter how good a treatment is prescribed, if congestion is not ingested, there is no meaning of treatment. Especially, dogs often have palatability. It is also common when it is not applicable. Therefore, there is a need to develop therapeutic agents that enhance palatability.

In recent years, the companion animal industry is continuously increasing, and accordingly, the importance of preventing the disease of companion animals is increasing. In particular, heart disease is increasing with the rapid increase of old dogs, and the need for treatment of heart disease such as chronic heart failure is increasing. In order to treat such heart disease, there is a need for a steady visit to an animal hospital and long-term treatment for heart disease.

According to the World Small Animal Heart Association (ISACHC) classification of heart failure, the disease class classified as Class I is The asymptomatic patient, which is not associated with clinical symptoms: cardiac murmur, arrhythmia (arrhythmia), or cardiac chamber enlargement (cardiac chamber enlargement) abnormality is confirmed, but the clinical symptoms, such as when there is no clear clinical symptoms of heart disease. The constriction classified as Class 1a is a case in which there is a symptom of heart disease but no symptom of a target action (volume or pressure overload ventricular hypertrophy). The axial axis classified as Class 1b is a case in which symptoms of heart disease are seen and the target action is detectable by ECG or radiation. Class II grafts, classified as Class II, show mild-to-moderate heart failure, and are clinical symptoms of heart disease that may affect the life of the graft in a resting state or with some exercise. If visible, typical clinical symptoms of cardiac disease, such as exercise avoidance, coughing, respiratory distress, and dyspnea, may be included. Hypoperfusion at rest may not be generally seen, and is a step in which hospital treatment is possible. Class III injuries classified as high grade heart disease (Advanced heart failure), when clinical symptoms of typical congestive heart failure can be clearly observed, dyspnea, dyspnea, ascites, severe exercise intolerance, etc. As a result, hypoperfusion at rest is also observed. When the condition becomes serious, symptoms of psychogenic shock may also be seen, and in this case, the prognosis may be poor. When classified as Class 3a, it is possible to go to a hospital (home care), while when classified as Class 3b, hospitalization is inevitable (psychic shock, life-threatening pulmonary edema, large amount of pleural effusion, etc.)

As a drug used for the treatment of these symptoms, ACE inhibitors, ARB agents, cardiac agents, nutritional supplements, thrombosis inhibitors, antiarrhythmic agents, etc. are applied, and for that treatment, a steady visit to an animal hospital and long-term supply of therapeutic agents are required. .

The above-mentioned background art is technical information acquired by the inventor for the derivation of the present invention or acquired in the derivation process of the present invention, and is not necessarily a known technology disclosed to the general public prior to the filing of the present invention.

Domestic patent application number, 10-1991-000822 U.S. Patent Application No., US 06/259537

An object of the present invention is to provide a preparation for the treatment or symptom relief of bicuspid valve degeneration in a canine animal, including an oral dosage form with improved palatability.

Another object of the present invention is to provide an oral administration preparation for canine animals with improved palatability.

Oral administration preparations for the treatment or symptom relief of bicuspid valve degeneration in canine animals according to an embodiment of the present invention include pimobendan or a pharmaceutically acceptable salt thereof; Pentoxifylline or a pharmaceutically acceptable salt thereof; And palatable excipients selected from the group consisting of beef powder, chicken liver powder and combinations thereof.

The pimobendan or a pharmaceutically acceptable salt thereof may be included as 0.005 to 0.02 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The oral dosage form may further include a sweetener.

The oral dosage form may further include an anti-caking agent.

The anti-caking agent may be selected from the group consisting of hydrous silicon dioxide, light anhydrous silicic acid, silicate hydrate, stearate, and combinations thereof.

The sweetener consists of aspartame, acesulfame potassium, sodium saccharin, sucralose, glycylic acid, stevia, sorbitol, erythritol, xylitol, maltitol, glucose, sucrose, fructose, xylitol, neohesperidine dihydrochalcone and combinations thereof. It may be selected from the group.

The palatable excipient may be contained in 1 to 20 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The sweetener may be contained in 0.1 to 1.5 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The oral dosage form may be administered as a powder, divided into 0.3 to 0.8 g / kgBW / day twice a day.

The oral dosage form may be added to the feed of canine animals to be fed.

The oral dosage form may be an improved rate of diastolic diameter of the left ventricle in a canine animal with bicuspid valve degeneration.

The oral dosage form may be to reduce the degree of coughing in canine animals with bicuspid valve degeneration.

Oral administration formulation for improved palatability for canine animals according to another embodiment of the present invention, pentoxifylline or a pharmaceutically acceptable salt thereof; And palatable excipients selected from the group consisting of beef powder, chicken liver powder and combinations thereof.

The oral dosage form may further include a sweetener.

The oral dosage form may further include an anti-caking agent.

The anti-caking agent may be selected from the group consisting of hydrous silicon dioxide, light anhydrous silicic acid, silicate hydrate, stearate, and combinations thereof.

The sweetener consists of aspartame, acesulfame potassium, sodium saccharin, sucralose, glycylic acid, stevia, sorbitol, erythritol, xylitol, maltitol, glucose, sucrose, fructose, xylitol, neohesperidine dihydrochalcone and combinations thereof. It may be selected from the group.

The palatable excipient may be contained in 1 to 20 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The sweetener may be contained in 0.1 to 3 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The pimobendan or a pharmaceutically acceptable salt thereof may be included in an amount of 0.005 to 0.02 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof.

The oral dosage form may be administered as a powder, divided into 0.3 to 0.8 g / kgBW / day twice a day.

The oral administration preparation may have a blood circulation improving activity including improving peripheral blood circulation and increasing coronary blood flow in the heart.

The oral dosage form may be added to the feed of canine animals to be fed.

The term "its pharmaceutically acceptable salt" or "salt thereof" described in connection with a specific compound in the present specification retains the biological effectiveness and properties of the parent compounds, and the single dose is When administered, it means salts that are not harmful biologically or in other directions. For example, "the pharmaceutically acceptable salt thereof" or "the salt thereof" may be a base addition salt of a specific compound, and may be prepared from inorganic and organic bases. Salts derived from inorganic bases can include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to primary, secondary and tertiary amines; Substituted amines, including naturally occurring substituted amines; And isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, Hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamine, theobromine, purine, piperazine, piperidine, and / or N-ethylpiperidine Salts of cyclic amines including dean. In addition, other carboxylic acid derivatives, such as carboxylic acid amides, including carboxamides, lower alkyl carboxamides, di (lower alkyl) carboxamides, etc., may also be useful in the practice of the present invention. have.

As used herein, the term "prevention" or "relaxation" refers to an act of suppressing progression, suppressing development, or alleviating the degree of occurrence of symptoms such as heart disease of a canine animal by administration of the agent of the present invention.

As used herein, the term "treatment" refers to the act of improving or beneficially altering symptoms such as heart disease of a canine animal by administering the formulation of the present invention, and means act of eliminating clinical symptoms or alleviating the degree of occurrence For example, the “treatment” may include relief or improvement of one or more symptoms or conditions among symptoms such as heart disease and medication stress, reduction of symptom range, stabilization of symptoms, non-proliferation of symptoms, suppression of symptoms, symptoms Delay or slow progression, improvement or alleviation of a symptom condition, and reduction of partial or overall symptom onset.

When describing the present invention in the present specification, no distinction is made between “singular” and “plural” of nouns, and singular expression is used as a concept including singular and plural, unless it is clear that the singular means only singular in context. do.

“Or”, “and / or” are used interchangeably, unless stated otherwise, and the description of “A or B” or “A and / or B” means “A, B, or A and B”. It can be interpreted as meaning including meaning.

Hereinafter, the present invention will be described in more detail.

To achieve the above object, the oral administration preparation for the treatment or symptom relief of bicuspid valve degeneration in a canine animal according to an embodiment of the present invention includes pimobendan or a pharmaceutically acceptable salt thereof; Pentoxifylline or a pharmaceutically acceptable salt thereof; And palatable excipients selected from the group consisting of beef powder, chicken liver powder and combinations thereof.

The pimobendan or a pharmaceutically acceptable salt thereof is a white or slightly yellow powder, which is not soluble in water but is soluble in dimethylformamide and slightly soluble in acetone and methanol. It has strong cardiac action and expands the arteries and veins. In addition, it has a phosphodiesterase III inhibitory action.

The pimobendan or a pharmaceutically acceptable salt thereof, along with the pentoxifylline or a pharmaceutically acceptable salt thereof, has an excellent effect when applied to treat or treat symptoms of bicuspid valve degeneration in a canine animal.

The pentoxifylline or a pharmaceutically acceptable salt thereof is a white or almost white crystalline powder that is soluble in water and soluble in methylene chloride, but not soluble in ethanol, and improves blood circulation activity. Have

The pimobendan or a pharmaceutically acceptable salt thereof may be included in an amount of 0.005 to 0.02 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof, and may be applied in an amount of 0.009 to 0.015 parts by weight.

The pentoxifylline or a pharmaceutically acceptable salt thereof has a strong bitter taste, and in particular, long-term feeding is difficult in reality, particularly because a mammal, such as a canine, has a clear preference for rejecting oral intake. Therefore, although long-term administration of a drug having a blood circulation improving activity such as pentoxifylline is necessary, in order to induce spontaneous oral intake of canine animals that refuse oral intake, a palatable excipient is applied in the present invention.

The palatability excipient refers to an excipient having a palatability of 80% or higher as suggested in the EU guideline. In the present invention, the palatability excipient is a beef powder or chicken liver powder that has been experimentally confirmed to be a palatability of 80% or higher. Includes. Preferably, chicken liver powder can be applied as the palatability excipient, which has been experimentally confirmed to have better palatability than the beef powder.

The oral dosage form may further include a sweetener.

The sweetener may improve the palatability of the oral dosage form by improving the palatability and the palatability of the bitter taste that may occur when the pentoxifylline is applied in large quantities.

As the sweetener, aspartame, acesulfame potassium, sodium saccharin, sucralose, glycylic acid, stevia, sorbitol, erythritol, xylitol, maltitol, glucose, sucrose, fructose, xylitol, neohesperidine dihydrochalcon, and combinations thereof It may be selected from the group consisting of, specifically, sodium saccharin, neohesperidin dihydrochalcone and a sweetener comprising a combination thereof may be applied.

The oral administration preparation having improved palatability for canine animals may further include an anti-caking agent.

The anti-caking agent (anticaking agent) may be any one selected from the group consisting of hydrous silicon dioxide, light anhydrous silicic acid, silicate hydrate, stearate, and combinations thereof.

The oral dosage form, the palatable excipient may be applied in an amount capable of sufficiently improving the palatability of the canine animal as a relative amount with the pentoxifylline or a pharmaceutically acceptable salt thereof, the pentoxifylline or its 1 part by weight of a pharmaceutically acceptable salt may contain 1 to 20 parts by weight of a palatable excipient, specifically 3 to 15 parts by weight of a palatable excipient, and more specifically 8 to 14 parts by weight of a palatable excipient may be included. . When a palatable excipient is included in such a content, palatability may be sufficiently improved upon oral intake of canine animals despite the bitter taste of pentoxifylline.

The sweetener serves to mask the bitter taste of the phenoxyphyllin and can also improve palatability when taken orally. Specifically, the sweetener is 0.1 based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof. It may be applied to 3 parts by weight, more specifically 1: 0.2 to 1.5 parts by weight, and 1: 0.4 to 0.8 parts by weight of sodium saccharin may be applied. When a sweetener is further included in such a content, palatability may be improved more than when only the palatability excipient is applied, and excellent palatability may be maintained even during continuous oral administration.

The anti-caking agent serves to help the components of the oral administration formulation containing the palatability excipient and the pentoxifylline to be sufficiently mixed and dispersed, for example, the pentoxifylline or a pharmaceutically acceptable salt thereof 1 It may be applied in an amount of 0.01 to 0.5 parts by weight based on parts by weight, and may be applied in an amount of 0.1 to 0.4 parts by weight.

The oral dosage form may further include additives such as carriers, other excipients, flavoring agents, dyes, and / or preservatives applied to pharmaceutical compositions other than those mentioned above.

In the case of the carrier or other excipient, it is applicable as a carrier or excipient not mentioned above, which is usually applied to veterinary preparations.

The flavoring agent may include fruit extracts (eg, fruit extracts such as vanilla, lemon, orange, grape, lime, apple, banana, pear, peach, strawberry, cherry, plum, pineapple, and apricot); One or more kinds may be selected from extracts of leaves, flowers, and fruits of plants. In addition, an artificial flavoring agent having an aroma such as an extract extracted from the fruit extract or the leaves of a plant may be used as the flavoring agent. In addition, flavoring agents such as artificial poultry scent may also be applied. The content of the flavoring agent may be applied in the range of 1 to 2% by weight relative to the total weight of the powder formulation.

The dye may be applied without limitation as long as it is a dye applied as a food additive or feed additive.

The oral administration preparation may be a powder, may be prepared by a method of mixing the respective compositions, and simple mixing by adding a pharmaceutically acceptable carrier such as lactose, starch, microcrystalline cellulose to the above-mentioned preparation Can also be produced.

The powder may have a particle size of 850 μm or less, and a particle size of 500 μm may be 95% by weight or more.

The oral dosage form may vary depending on the content of the active ingredient contained in the preparation, the condition of the individual (weight, etc.), the symptoms and side effects being treated, or the severity of the stress state, etc. It may be divided into several times and administered several times during the day, specifically, 0.3 to 0.8 g / kgBW / day may be administered twice a day, and 0.35 to 0.55 g / kgBW / day may be administered twice a day. Can be.

The oral dosage form may further include a dehumidifying agent that is packaged in a water-impermeable packaging material and does not directly contact the preparation inside the packaging.

The oral dosage form, including pimobendan or a pharmaceutically acceptable salt thereof, and pentoxifylline or a pharmaceutically acceptable salt thereof, may have improved cardiac effects and improved blood circulation activity.

In addition, in spite of the bitter taste derived from at least some of the pharmacologically active substances, palatability was significantly improved to enable spontaneous oral administration to canine animals, thereby significantly improving intakeability during long-term feeding.

The oral dosage form may have improved the rate of improvement of the left ventricular diastolic diameter of canine animals with bicuspid valve degeneration, which was able to experimentally confirm a significant effect compared to when pimobendan was applied alone.

The oral administration formulation may be to reduce the degree of coughing in canine animals having bicuspid valve degeneration, which can be experimentally confirmed as a significant effect compared to when pimobendan was applied alone.

In addition, the oral dosage form has a circulating personality activity such as improved peripheral blood circulation and increased coronary blood flow in the heart, and helps the canine animal with bicuspid valve degeneration to have a good prognosis.

Oral administration preparations for the treatment or symptom relief of bicuspid valve degeneration in the canine animals, in addition to the above-mentioned drugs, diuretics, ACE inhibitors, ARB preparations, cardiac and nutritional supplements, thrombosis inhibitors, antiarrhythmic agents, fatty acids, etc., are added as necessary. May be

As the diuretic, furosemide, torcemide, spironolactone, hydrochlorothiazide, and the like can be applied. As the ACE inhibitor, enalapril, ramipril, benazapril, and the like, and as an arterial expansion agent, hydralazine , Amlodipine as a calcium channel blocker, sildenafil, tadalafil as a pulmonary arterial expander, nitrovascular extender as an arteriovenous extender, digoxin, catecholamine, etc. as the strong core agent.

The oral dosage form can be applied as a pharmaceutical composition.

The oral dosage form may be included in the food composition for canine animals.

The oral dosage form may be applied as a feed additive composition.

The oral dosage form may be applied as a jerky additive composition.

The oral dosage form may be provided so that the canine can be voluntarily taken in a manner of oral administration, and may be fed separately from or added to the feed. In this case, the palatability of the oral administration preparation is significantly improved, so that both the administering agent and the ingesting canine animal are relieved of stress on feeding / ingestion, and long-term administration of the necessary drug is possible through oral ingestion.

In order to achieve the above object, an oral administration preparation with improved palatability for a canine animal according to another embodiment of the present invention, pentoxifylline or a pharmaceutically acceptable salt thereof; And palatable excipients selected from the group consisting of beef powder, chicken liver powder and combinations thereof.

The pentoxifylline or a pharmaceutically acceptable salt thereof is a white or almost white crystalline powder that is soluble in water and soluble in methylene chloride, but not soluble in ethanol, and improves blood circulation activity. To have. This has a strong bitter taste, and in particular, long-term feeding is difficult in reality because mammals, which are clearly palatable, such as canine animals, often refuse oral intake. Therefore, although long-term administration of a drug having a blood circulation improving activity such as pentoxifylline is necessary, in order to induce spontaneous oral intake of canine animals that refuse oral intake, a palatable excipient is applied in the present invention.

The palatability excipient refers to an excipient having a palatability of 80% or higher as suggested in the EU guideline. In the present invention, the palatability excipient is a beef powder or chicken liver powder that has been experimentally confirmed to be a palatability of 80% or higher. Includes. Preferably, chicken liver powder can be applied as the palatability excipient, which has been experimentally confirmed to have better palatability than the beef powder.

The oral dosage form may further include a sweetener.

The sweetener may further improve the palatability of the oral administration formulation while simultaneously shielding the bitter taste that may occur when the pentoxifylline is applied in a large amount.

As the sweetener, aspartame, acesulfame potassium, sodium saccharin, sucralose, glycylic acid, stevia, sorbitol, erythritol, xylitol, maltitol, glucose, sucrose, fructose, xylitol, neohesperidine dihydrochalcon, and combinations thereof It may be selected from the group consisting of, specifically, sodium saccharin, neohesperidin dihydrochalcone and a sweetener comprising a combination thereof may be applied.

The oral administration preparation having improved palatability for canine animals may further include an anti-caking agent.

The anti-caking agent (anticaking agent) may be any one selected from the group consisting of hydrous silicon dioxide, light anhydrous silicic acid, silicate hydrate, stearate, and combinations thereof.

The oral dosage form, the palatable excipient may be applied in an amount capable of sufficiently improving the palatability of the canine animal as a relative amount with the pentoxifylline or a pharmaceutically acceptable salt thereof, the pentoxifylline or its 1 part by weight of a pharmaceutically acceptable salt may contain 1 to 20 parts by weight of a palatable excipient, specifically 3 to 15 parts by weight of a palatable excipient, and more specifically 8 to 14 parts by weight of a palatable excipient may be included. . When a palatable excipient is included in such a content, palatability may be sufficiently improved upon oral intake of canine animals despite the bitter taste of pentoxifylline.

The sweetener serves to mask the bitter taste of the pentoxifylline and can improve palatability when taken orally. Specifically, the sweetener is 0.1 to 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof. It may be applied to 3 parts by weight, more specifically 1: 0.2 to 1.5 parts by weight, and 1: 0.4 to 0.8 parts by weight of sodium saccharin may be applied. When a sweetener is further included in such a content, palatability may be improved more than when only the palatability excipient is applied, and excellent palatability may be maintained even during continuous oral administration.

The anti-caking agent serves to help the components of the oral administration formulation containing the palatability excipient and the pentoxifylline to be sufficiently mixed and dispersed, for example, the pentoxifylline or a pharmaceutically acceptable salt thereof 1 It may be applied in an amount of 0.01 to 0.5 parts by weight based on parts by weight, and may be applied in an amount of 0.1 to 0.4 parts by weight.

The oral dosage form may further include additives such as carriers, excipients, flavoring agents, dyes, and / or preservatives applied to pharmaceutical compositions other than those mentioned above.

The carrier or excipient may be applied as long as it is a carrier or excipient that is usually applied to veterinary preparations.

The flavoring agent may include fruit extracts (eg, fruit extracts such as vanilla, lemon, orange, grape, lime, apple, banana, pear, peach, strawberry, cherry, plum, pineapple, and apricot); One or more kinds may be selected from extracts of leaves, flowers, and fruits of plants. The content of the flavoring agent may be applied in the range of 1 to 2% by weight relative to the total weight of the powder formulation.

The dye may be applied without limitation as long as it is a dye applied as a food additive or feed additive. For example, a pigment such as Red No. 40 can be used in a trace amount (for example, about 0.01% by weight).

The oral dosage form may be a powder, may be prepared by a method of mixing the composition, or may be prepared by additionally simple mixing of a suitable pharmaceutically acceptable carrier such as lactose, starch or microcrystalline cellulose.

The powder may have a particle size of 850 μm or less, and a particle size of 500 μm may be 95% by weight or more.

The oral dosage form may vary depending on the content of the active ingredient contained in the preparation, the condition of the individual (weight, etc.), the symptoms and side effects being treated, or the severity of the stress state, etc. It may be divided into several times and administered several times during the day, specifically, 0.3 to 0.8 g / kgBW / day may be administered twice a day, and 0.35 to 0.55 g / kgBW / day may be administered twice a day. Can be.

The oral dosage form may further include a dehumidifying agent that is packaged in a water-impermeable packaging material and does not directly contact the preparation inside the packaging.

The oral dosage form, including pentoxifylline having pharmacological activity or a pharmaceutically acceptable salt thereof, has a blood circulation improving activity, and spontaneous oral administration to a canine animal is possible despite the bitter taste of the pharmacologically active substance. To improve the palatability so much.

The oral dosage form has a blood circulation personality activity, such as improved peripheral blood circulation and increased coronary blood flow in the heart, and can be administered in combination with other drugs as necessary. For example, the oral administration preparation, in addition to the above-mentioned drugs, diuretics, ACE inhibitors, ARB preparations, cardiac and nutritional supplements, thrombus inhibitors, antiarrhythmic agents, fatty acids, etc. may be added as needed. As the diuretic, furosemide, torcemide, spironolactone, hydrochlorothiazide, and the like can be applied. As the ACE inhibitor, enalapril, ramipril, benazapril, and the like, and as an arterial expansion agent, hydralazine , Amlodipine as a calcium channel blocker, sildenafil, tadalafil as a pulmonary arterial expander, nitrovascular extender as an arteriovenous extender, digoxin, catecholamine, etc. as the strong core agent.

The oral dosage form can be applied as a pharmaceutical composition.

The oral dosage form may be included in the food composition for canine animals.

The oral dosage form may be applied as a feed additive composition.

The oral dosage form may be applied as a jerky additive composition.

The oral dosage form may be provided so that the canine can be voluntarily taken in a manner of oral administration, and may be fed separately from or added to the feed. In this case, the palatability of the oral administration preparation is significantly improved, so that both the administering agent and the ingesting canine animal are relieved of stress on feeding / ingestion, and long-term administration of the necessary drug is possible through oral ingestion.

The oral administration preparations with improved palatability of the present invention have excellent palatability, which facilitates long-term oral administration to dogs and animals, and can be applied as a powder or the like with pimobendan to provide a formulation for the treatment of heart disease or symptom relief of dogs. have.

1 is a graph of changes in pimobendan concentration over time in an experimental group through LC-MS / MS among pharmacokinetic test results conducted in an embodiment of the present invention.
Figure 2 is a graph of the change in the concentration of pentoxifylline with time of the experimental group through LC-MS / MS among the results of pharmacokinetic tests conducted in the Examples of the present invention.
Figure 3 is a graph showing the results of observing the change over time of the cardiovascular index FS (fractional shortening), the pharmacodynamics test results conducted in an embodiment of the present invention (* is the corresponding time based on when time = 0 Significant difference in (P <0.05), † means significant difference (P <0.05) based on the result of the comparative example at the same time).
Figure 4 is a graph showing the result of observing the change over time of the cardiovascular indicator LVIDs (left ventricular internal diameter, systole) of the pharmacodynamics test results in the embodiment of the present invention (* is based on the time = 0 The significant difference (P <0.05) at the corresponding time, and † means the significant difference (P <0.05) based on the result of the comparative example at the same time).
Figure 5 is a graph showing the results of observing the change over time of the cardiovascular indicator PEP (pre-ejection period) of the results of the pharmacodynamics test conducted in the embodiment of the present invention (* is based on the time = 0 Significant difference (P <0.05) at the time, † means significant difference (P <0.05) based on the results of the comparative examples at the same time).
6 is an ECG test result of group 1 in the safety experiment conducted in the embodiment of the present invention.
7 is a group 2 ECG test results in the safety experiment conducted in the embodiment of the present invention.
8 is an ECG test result of group 3 in the safety experiment conducted in the embodiment of the present invention.
9 is an ECG test result of group 4 in the safety experiment conducted in the embodiment of the present invention.
Figure 10 is a side examination of the heart and kidneys of the experimental dog of Group 4 in the safety experiment conducted in the embodiment of the present invention.
11 is a photograph of the results of a cardiac biopsy performed in a safety experiment conducted in an embodiment of the present invention.
12 is a result of chest radiograph taken in a small-scale short-term evaluation of the present invention.
13 is an echocardiographic evaluation picture of a control group conducted in a small-scale short-term evaluation of the present invention.
14 is an echocardiographic evaluation picture of an embodiment conducted in a small-scale short-term evaluation of the present invention.
15 is an ECG evaluation result of the control group conducted in the small-scale short-term evaluation of the present invention.
16 is an ECG evaluation result of the experimental group conducted in the small-scale short-term evaluation of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art to which the present invention pertains can easily practice. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein.

1. Test and evaluation of excipients to enhance palatability

(1) Experiment method

This experiment was conducted with the approval of the Laboratory Animal Ethics Committee (IACUC).

-Experiment preparation

Healthy dogs of various conditions were selected as experimental animals, regardless of breed and sex.

The candidate excipients include processed seafood (eg bonito extract, oyster extract, katsuo concentrate powder, anchovy powder, cod roe powder, shrimp powder, shrimp extract, tuna powder, abalone powder), and livestock processed products (beef extract powder, beef powder, chicken powder) , Chicken liver powder), and agricultural products processed products (ginger powder, garlic powder, pepper powder, cone granules), among which substances are expected to improve palatability, after limiting the scope of the candidate group, among them, beef powder, chicken liver Powder and cheonggukjang were selected as candidates for excipients.

-Setting of test method

A preliminary investigation into the palatability test of veterinary medicines was conducted, and the test method was designed based on the paper on palatability testing of companion animal medicinal products published in South Africa in 2014 and the palatability test guidelines prescribed by EMA. . Also, the study conducted in 2007 was used for statistical analysis.

Reference 1: Comparison of the Palatability of a New Flavored Drontal® Plus Tablet (Drontal® Plus Treat 10 kg) and Milbemax® Chewable Tablets When Presented to Privately Owned Dogs, Open Journal of Veterinary Medicine, 2014, 4, 163-169, Gabriele Petry, Josephus Fourie, Sonja Wolken

Reference 2: Guideline on the demonstration of palatability of veterinary medicinal products EMA / CVMP / EWP / 206024 / 2011-CONSULTATION

Reference 3: An evaluation of the relative palatability of two commercial oral tablet formulations of carprofen and meloxicam in dogs using acceptance and preference tests, Revue Med. Vet., 2007, 158, 10, 519-524, M. Payne-Johnson, T. P. Maitland, N. Tilt and J. Gossellin

From April 8, 2017 to May 2, 2107, an experiment was conducted on a farm in Yongin-si, Gyeonggi-do, and divided into 3 groups of 10 dogs each for healthy dogs of various conditions regardless of breed and gender. A palatability test was conducted on. In order to exclude variables such as specific conditions, the experiment was conducted without changing the experiment location, feed type, and payer.

30 healthy experimental dogs were divided into 3 groups of 10 each, and beef powder, chicken liver powder, and cheonggukjang powder were fed every 3 days, and the amount of spontaneous feeding was observed, and the results were summarized. The results were checked by dividing them into three items: 'voluntarily eating all the amounts', 'partially eating within a given time', or 'rejecting voluntary intake', and the same experiment was repeated three times.

The experimental results were calculated as follows, and statistical significance was evaluated by calculating the chi-square.

Palatability success (%) (when consumed voluntarily within 1 minute)

= (Number of experiment dogs that voluntarily ate all / number of experiment dogs) x 100

(2) Test results

-Determination of the amount of benefits

Prior to this experiment of palatability test, an excipient feeding amount determination experiment was conducted. 10 animals are randomly selected, and for convenience of feeding, 10 g chicken liver powder, which is 1 tbsp standard, and 2 g chicken liver powder, which is 1 tsp standard, are given, and all test dogs eat all the excipients given within a given time. The possible amount was evaluated. As a result of performing the repeated test twice in the same experiment, 2 g of the excipient, in which all 20 excipients were able to eat all the excipients in a short time, was determined as the dose of the palatability test.

-Evaluation of palatability

During the first experiment, test dogs from group A showed interest in beef powder as a whole, but three did not fully consume the given powder within a given minute, and the test dogs from group B who received cheonggukjang powder were mostly consumed. Refused or ate a little. All C group test dogs who received chicken liver powder ate all the chicken liver powder given within a given time.

When conducting the second experiment, most of the group A dogs who received the Cheonggukjang powder remained, and the group B dogs who received chicken liver powder all ate well, and the group C dogs who received beef were also partially consumed. All but the test dog of the dog ate well.

When conducting the 3rd experiment, all of the test dogs except group 2, which received chicken liver powder, ate chicken liver powder well, and group B, which received beef powder, also ate all the excipients given by the test dogs. However, most of the C group's test dogs that received Cheonggukjang powder only smelled and immediately lost interest.

These results showed a similar tendency for the second retest and the third retest, and the above results are summarized in Table 1 below.

Beef powder Chicken liver powder Cheonggukjang powder Entire Partial Refusal Entire Partial Refusal Entire Partial Refusal Primary April 08 7 3 0 10 0 0 0 4 6 April 11 8 2 0 10 0 0 0 One 9 April 14 9 One 0 8 2 0 0 7 3 Secondary April 17 8 2 0 9 One 0 0 6 4 April 20 8 2 0 9 One 0 0 5 5 April 23 8 2 0 9 One 0 0 5 5 3rd April 26 9 One 0 8 2 0 0 7 3 April 29 8 2 0 9 One 0 0 7 3 May 02 7 3 0 9 One 0 0 4 6 Total 72 18 0 81 9 0 0 46 44 Palatability success (%) 80 20 0 90 10 0 0 51.11 48.89

Referring to the results in Table 1 above, the calculated palatability success rate indicated that both the beef powder and chicken liver powder exceeded the success criteria (80% beef powder, 90% chicken liver powder). However, even though the number of times it was estimated to be partially consumed was similar, there was a significant difference in the remaining amount. Most of the test dogs did not consume all of them within 1 minute, but the amount of chicken liver powder left was small, compared with the number of times that most of them left 1/2 to 1/4 of the initial amount. There was a turn. Therefore, it was evaluated that the palatability of the chicken liver powder was much higher than that of the beef powder when the test dogs considered the amount left after the end of the test.

In the following tests, chicken liver powder was applied as a palatable excipient.

2. Preparation of pharmaceutical composition with improved palatability and pharmacodynamic evaluation

(1) palatability test

-Composition of Example 1 and palatability evaluation results

Based on the palatability test results conducted in April to May 2017, the composition of Example 1 for the preparation of a mixture of pentoxifylline and pimobendan with improved palatability by adding chicken liver powder was set as shown in Table 2 below.

ingredient Pimobendan Pentoxifylline Anti-caking agent
(Silica powder) Excipient
(Corn starch) Palatable excipient
(Chicken liver powder) content 10.102 mg 0.594 g 0.132 g 2.970 g 2.894 g

Using the prototype according to the composition of Example 1, the palatability of the test dogs for safety testing was investigated. As a result, there was no problem in ingestion when feeding a prototype with a dose of 4 times, but there were test dogs who refused to ingest when palatability was investigated to overdose 16 times the dose. As a cause of this, the bitter taste of pentoxifylline was strong and it was determined that the content of chicken liver powder in the existing palatability test was insufficient to enhance palatability.

-Composition of Example 2 and results of palatability evaluation

The composition to increase the content of chicken liver powder was calculated again, and a prototype was prepared with the composition shown in Table 3 below.

ingredient Pimobendan Pentoxifylline Anti-caking agent
(Silica
powder) Sweetener
(saccharin
salt) Palatability
Excipient
(Chicken liver powder) total content
(Parts by weight) 1.25 75 20 50 853.75 1,000

As a result of conducting a palatability test with the composition of Example 2, it was confirmed that, unlike the composition of Example 1, there was no problem of palatability even in the case of overdose of 16 times as well as the quantitative and 4 times dose. Therefore, in the following experiment, the composition of Example 2 was applied.

(2) Pharmacodynamics test (PD)

-Experimental materials and methods

Eight healthy experimental dogs (beagle dogs) were used as healthy individuals without other diseases, and a randomized, experimenter-blind single dose, same dose experiment method was conducted.

Experimental dogs were randomly divided into two groups, the experimental group and the control group (4 each), and on the day of drug administration, the oral dose of each group was administered once. The control group was fed with 10 mL of water, and the experimental group was orally administered with Pimobendan 0.27 mg / kg (range: 0.26-0.28 mg / kg) and Pentoxifylline 15 mg / kg using the composition of Example 2. All subjects were fed 5-10 mL of water after administration to ensure drug intake.

-Pharmacokinetic analysis

Pharmacokinetics analysis was conducted by requesting an external laboratory, and after pre-treatment of serum separated from blood collected from experimental dogs after ingestion of the drug and after a certain time, pimobendan and pentoxifylline were obtained by LC-MS / MS. PK analysis was performed by analyzing the content of.

-Echocardiography and blood pressure measurement

Experimental dogs were pre-trained to become familiar with the Lateral recumbency posture before conducting echocardiography. No other sedative drugs were used other than the corrector. Echocardiography was operated by a single investigator during this experiment, and an EPIQ 7 Cardiology Ultrasound Machine (Philips, USA) was used.

Survey items were: LA size (mm, left atrial size), Ao size (mm, aortic size, inner diameter), LA / Ao (left atrium to aortic ratio), IVSd (mm, diastolic ventricular septal thickness), IVSs (mm, contraction) End ventricular septal thickness), LVIDd (mm, diastolic left ventricular diameter), LVIDs (mm, systolic left ventricular diameter), LVPWd (mm, diastolic left ventricular wall thickness), LVPWs (mm, systolic left ventricular posterior wall thickness), FS ( %, Left ventricular shortening rate), Ao vel (maximum aortic blood flow rate), PEP (pre-contraction period), and LVET (left ventricular contraction time).

-Pharmacodynamics analysis

The size of IVSs, IVSDd, LVIDs, LVIDd, LVPWs, LVPWd, LA, and Ao can be affected depending on the size of the dog (Cornell et al., 2004), so the values were corrected through weight. The difference in indicators between the two groups was judged to be significant when the p-value was 0.05 or less. The results of the pharmacodynamic analysis are expressed as the mean and standard deviation.

(3) Pharmacodynamics test results

-Analysis of experimental results

In the analysis of pharmacokinetics, at least three tests were performed for each concentration using a QC sample to calculate accuracy (recovery rate) and precision (variation coefficient, CV).

A calibration curve was prepared to quantify pimobendan and pentoxifylline in serum, the detection limit (LOD) was 3 times the S / N (signal to noise) ratio, and the quantitation limit (LOQ) was at least 10 times the S / N ratio It was calculated as. For mass spectrometry, API 4000 MS / MS System (SCIEX, USA) was used, and positive ions fragmented by ESP (Electrospray Ionization) were detected in MS / MS mode. LC-MS / MS was used to measure pimobendan and pentoxifylline blood peak concentration time (Tmax), blood peak concentration (Cmax), blood distribution volume (AUC (inf)) and half-life (T1 / 2). .

In Pharmacodynamics analysis, all data are presented as mean ± standard deviation. The Mann-Whitney test method was used to compare data between the experimental group and the control group and to compare the data before and after drug administration. LA size, Ao size, LA / Ao, IVSd, IVSs, LVIDd, LVIDs, LVPWd, LVPWs, FS, Ao vel, PEP, LVET and systolic blood pressure, diastolic blood pressure, and mean arterial pressure were measured over time after drug administration. It was analyzed whether the difference between the experimental group and the control group shown in, or before and after drug administration had statistical significance. In the case of P-value 0.05 or less, it was judged to be significant. SPSS version 20 (SPSS, Inc., Chicago, IL, USA) was used for statistical analysis.

-Pharmacokinetics test results

The change in concentration of pimobendan over time in the experimental group after oral administration of pimobendan is shown in FIG. 1, and the change in concentration of pentoxifylline with time in the experimental group after oral administration of pentoxifyl is shown in FIG. 2.

1 and 2, as a result of analyzing the pharmacokinetics of pimobendan using LC-MS / MS, the peak concentration of pimobendan in all subjects (Tmax) was 1.08 ± 0.72 hours, and the highest in blood. Concentration (Cmax) was found to be 2.83 ± 1.50 ng / mL. The volume of blood distribution (AUC (inf)) was 5.04 ± 0.84, and the half-life (T1 / 2) was 2.65 ± 1.42 hours. As a result of analyzing the pharmacokinetics of pentoxifylline, the peak concentration of blood (Tmax) of pentoxifylline in all subjects was 0.29 ± 0.14 hours, and the highest concentration in blood (Cmax) was 1184.33 ± 932.37 ng / mL. The volume of blood distribution (AUC (inf)) was 1648.15 ± 723.56, and the half-life (T1 / 2) was 0.29 ± 0.23 hours.

-Pharmacodynamics test results

As a result of the basic echocardiography test before the experiment, it was confirmed as shown in Table 4 below that there was no significant difference between the control group and the experimental group. Table 4 below shows the basic echocardiographic index of the control group (Control; n = 4) administered only with water before the experiment (before drug administration) and the experimental group administered with the composition of Example 2 (PP; n = 4).

Parameter (units) Control Experiment group (PP) Ao Vel (m / s) 127.4 ± 30.2 123.3 ± 29.8 Ao * (cm) 1.7 ± 0.0 1.5 ± 0.6 FS (%) 43.1 ± 4.3 37.7 ± 2.5 HR (bpm) 134.5 ± 16.0 94.3 ± 17.9 IVSd * (cm) 0.9 ± 0.1 0.8 ± 0.1 IVSs * (cm) 1.2 ± 0.2 1.1 ± 0.1 LA * (cm) 2.0 ± 0.1 1.8 ± 0.7 LVET (ms) 174.2 ± 7.1 198.7 ± 9.2 LVIDd * (cm) 3.0 ± 0.2 2.6 ± 0.8 LVIDs * (cm) 1.7 ± 0.2 1.6 ± 0.5 LVPWd * (cm) 0.9 ± 0.2 0.7 ± 0.2 LVPWs * (cm) 1.3 ± 0.1 1.1 ± 0.2 PEP (ms) 20.5 ± 3.1 25.8 ± 4.6 MBP (mmHg) 94.3 ± 6.7 101.5 ± 8.6 DBP (mmHg) 74.5 ± 13.5 90.0 ± 7.5 SBP (mmHg) 127.5 ± 10.1 123.5 ± 9.9 Ao Vel, aortic velocity; Ao, aortic diameter; FS, fractional shortening; HR, heart rate; IVSd, interventricular septum (diastole); IVSs, interventricular septum (systole); LA, left atrium; LVET, left ventricular ejection time; LVIDd, left ventricular internal diameter (diastole); LVIDs, left ventricular internal diameter (systole); LVPWd, left ventricular posterior wall (diastole); LVPWs, left ventricular posterior wall (systole); PEP, pre-ejection period; MBP, mean blood pressure; DBP, diastolic blood pressure; SBP, systolic blood pressure.
No values were significantly different between treatment groups (P> 0.05).
Values expressed as mean standard ± deviation.
* Indexed to body weight (Cornell et al., 2004).

In the control group (Control; n = 4) administered only with water and in the experimental group (PP; n = 4) administered pimobendan and pentoxifylline, changes in cardiovascular indicators were observed over time, and changes in FS, LVIDs, and PEP were respectively observed. 3 to 5 are shown.

As a result of the experiment, the indicators showing significant differences from the existing indicator values were as follows: FS (1-4 h), LVIDs (1-3 h), PEP (3, 4 h), FS increased compared to the existing indicator values. LVIDs and PEP decreased. The maximum effect of the drug appeared between 2 hours and 4 hours after administration, and the significant difference (P <0.05) between the experimental group and the control group was 2 to 4 hours in FS and 1 to 3 hours in LVIDs. , And at 3 and 4 hours in PEP.

Taken together, the composition of Example 2 above, which is a combination of pimobendan and pentoxifylline, could be appropriately administered through oral administration, with a half-life (T _1/2 ) of pimobendan 2.65 ± 1.42 hours and pentoxifylline 0.29 ± 0.23 hours. Was quickly discharged from the body. The maximum time to reach blood concentration (Tmax) in all subjects was pimobendan 1.08 ± 0.72 hours, pentoxifylline 0.29 ± 0.14 hours, and the highest blood concentration (Cmax) was pimobendan 2.83 ± 1.50 ng / mL, pentoxifylline 1184.33 ± It was 932.37 ng / mL. Blood distribution volume (AUC (inf)) was pimobendan 5.04 ± 0.84, pentoxifylline 1648.15 ± 723.56.

As a result of the single administration in the above example, it was confirmed through echocardiography that the composition significantly enhanced the contractile function of the heart. The fractional left ventricular shortening increased over time, and showed significant differences compared to the pre-drug values at 1 hour, 1.5 hours, 2 hours, 3 hours, and 4 hours after drug administration. Significant differences were shown at 2, 3 and 4 hours after drug administration compared to the control group. Left ventricular internal diameter at end systole showed a decreasing pattern with time, and it was significant compared to the existing values before drug administration at 1 hour, 1.5 hours, 2 hours, and 3 hours after drug administration. Differences were shown, and 1 hour, 1.5 hours, 2 hours, and 3 hours after drug administration compared to the control group. The pre-ejection period showed a decreasing pattern with time, and showed significant differences compared to the pre-drug values at 3 and 4 hours after drug administration. Changes in these indicators were used as useful indicators for confirming the effect of the drug, and it was also possible to confirm the duration of the drug effect. The maximum effect of the drug was observed 2-4 hours after administration, and the effect was maintained for a time within 8 hours.

3. Safety evaluation

(1) Experiment method

Toxicity was administered at various doses in single and repeated doses.

Ten clinically healthy beagle dogs, regardless of sex, were used in the experiment. The body weight is around 8-14kg, and all subjects undergo a medical examination including an overall physical examination before the start of the experiment, and then have an adjustment period in the same experimental breeding facility for 1 week. For the feed, dry feed of the same brand was fed twice a day at a fixed time, and water was allowed to be fed at all times. The breeding facility where the experimental animals stayed was intended to maintain a constant temperature, humidity, and artificial illuminance. Both animal breeding and management were approved by the Animal Experimental Ethics Committee and conducted in accordance with the regulations.

The experimental dogs were randomly divided into 4 groups. Each group was divided into Group 1 (control group), Group 2 (recommended dose-administered group), Group 3 (recommended 4-fold overdose group), and Group 4 (recommended 16-fold overdose group), and Group 2 (recommended group). Dose group) was assigned to 4 animals, and the remaining group 1, group 3 and group 4 were assigned to 2 animals per group. Different doses of drugs were orally administered for a total of 4 weeks, and the doses of drugs administered to each group are shown in Table 5 below.

group Number of objects Drug dosage Toxic dose Method of drug administration Pimobendan Pentoxifylline One 2 horses 0 mg / kg / day 0 mg / kg / day Control No drug administration 2 4 horses 0.5 mg / kg / day 30 mg / kg / day Recommended dosage Divided twice a day for a total of 4 weeks 3 2 horses 2.0 mg / kg / day 120 mg / kg / day 4 times the recommended dose 4 2 horses 8.0 mg / kg / day 480 mg / kg / day 16 times the recommended dose

(2) Evaluation of safety

- Assessment Methods

Mortality / Morbidity was evaluated by recording the mortality or morbidity that occurred by observing the test dog every day for a period of 4 weeks.

In the Physical Examination section, daily abnormalities are checked on a basic physical examination for a period of 4 weeks, and feed intake is checked to check for health abnormalities. Body weights were measured on days 0, 7, 14, 21 and 28.

Blood examination includes: whole blood count (CBC; WBC, RBC, Hb, HCT, MCV, MCH, MCHC, PLT), serochemistry (BUN, Cre, ALT, AST, ALP, TP, Alb) And electrolyte tests (Na, K, Cl) were performed on day 0, day 7, day 14, day 21, and day 28 to check for abnormality.

Urinalysis was performed on the 0th, 7th, 14th, 21st, and 28th days to evaluate urine function according to drug administration. Urine specific gravity (USG), Urine dip stick, Wet preparation and cytology were performed.

For electrocardiography (Electrocardiography), electrocardiography is performed on Days 0, 7, 14, 21, and 28 to check for abnormalities such as arrhythmia, atrial fibrillation, and other abnormal waveforms due to drug administration. ECG-1350 (Nihon kohden, Tokyo, Japan) was used as the ECG device, and the beagle dog was measured in a Lateral recumbency or sitting or standing position. No sedative drugs were used other than the corrector.

In echocardiography, echocardiography is performed on Days 0 and 28 to check the morphological changes and basic functional indicators of the heart and check for abnormalities. EPIQ 7 Cardiology Ultrasound Machine (Philips, USA) was used as the echocardiogram device.

The necropsy was visually evaluated for organ (cardiac, lung, liver, spleen, kidney, and digestive) toxicities through an autopsy after the 28-day experiment, and the heart was examined for histopathological changes through biopsy. Did.

- Evaluation results

As a result of mortality / morbidity evaluation, daily observation dogs were observed during the four-week experimental period, and no death or disease-related experimental dogs were observed, including dysfunction, irritability, and digestive side effects. No drug side effects were observed.

As a result of the physical examination, the results of changes in the physical examination results according to the measurement timing during the experiment are shown in Table 6. No significant changes were observed in the body weight, heart rate, respiratory rate and body temperature during the experiment.

Variables Day 0 Day 7 Day 14 Day 21 Day 28 BW (Kg) 10.5 ± 2.3 10.4 ± 2.1 10.5 ± 1.9 10.6 ± 1.7 10.5 ± 2.1 HR (bpm) 116 ± 24 121 ± 9 127 ± 10 114 ± 11 112 ± 8 RR (/ min) 32 ± 4 34 ± 3 28 ± 6 24 ± 2 32 ± 3 BT (℃) 38.4 ± 0.4 38.7 ± 0.4 38.5 ± 0.8 38.6 ± 0.5 38.5 ± 0.4 * BW, body weight; HR, heart rate; RR, respiratory rate; BT, body temperature.

The results of blood examination (Blood examination) were observed in Table 7 below by observing the change in blood test results according to the measurement time point during the experiment. During the experimental period, no significant changes were observed in the subject's whole blood cell test, serum chemistry, and electrolyte test. All values were within the reference range.

Variables Day 0 Day 7 Day 14 Day 21 Day 28 Reference value CBC WBC
(x10 ³ / ul) 10.1
± 2.1 9.5
± 2.1 9.1
± 2.1 9.3
± 1.5 10.0
± 1.7 5.2 ~ 13.9 RBC
(x10 ⁶ / ul) 7.3
± 1.0 7.3
± 0.7 6.7
± 0.9 7.4
± 1.0 7.3
± 0.9 5.7 ~ 8.8 Hb (gm / dl) 16.4
± 2.4 16.5
± 1.5 15.0
± 1.8 16.5
± 1.8 16.2
± 1.7 12.9 ~ 18.4 HCT (%) 52.2
± 9.1 51.5
± 4.7 46.5
± 6.7 52.0
± 6.2 51.0
± 6.9 37.1 ~ 57.0 MCV (fL) 71.1
± 2.2 70.3
± 1.9 63.4
± 20.2 70.4
± 2.5 70.1
± 3.3 58.8 ~ 71.2 MCH (pg) 20.6
± 6.4 22.6
± 0.8 20.4
± 6.3 22.4
± 0.8 22.3
± 0.6 20.5 ~ 24.2 MCHC (%) 31.7
± 1.3 32.1
± 5.5 32.3
± 1.5 31.8
± 0.9 31.8
± 1.3 31.0 ~ 36.2 PLT (x10 ³ / ul) 281
± 69 284
± 76 255
± 69 296
± 74 240
± 73 143 ~ 400 Serum Chemistry TP
(g / dL) 6.9
± 0.4 6.8
± 0.5 6.2
± 0.5 6.2
± 0.5 6.2
± 0.6 5.4 ~ 7.4 ALB
(g / dL) 3.3
± 0.1 3.4
± 0.2 3.2
± 0.1 3.3
± 0.2 3.2
± 0.1 2.9 ~ 4.2 BUN (mg / dL) 14.2
± 3.3 10.6
± 3.3 10.6
± 3.8 12.2
± 4.3 13.3
± 4.8 8 ~ 26 CREA (mg / dL) 0.7
± 0.1 0.8
± 0.1 0.7
± 0.1 0.8
± 0.1 0.9
± 0.1 0.5 ~ 1.3 AST (U / L) 33.7
± 7.4 33.2
± 5.9 26.1
± 5.3 25.5
± 5.0 25.6
± 4.7 15 ~ 43 ALT (U / L) 34.9
± 14.0 33.9
± 12.6 38.4
± 15.8 34.7
± 13.5 35.0
± 21A.0 19 ~ 70 ALP (U / L) 55.8
± 28.3 47.3
± 22.0 47.3
± 21.5 50.4
± 21.4 58.6
± 24.9 15-127 Na (mmol / L) 145.9
± 1.9 145.1
± 0.6 145.7
± 2.0 145.6
± 1.6 146.5
± 2.3 144 ~ 154 K (mmol / L) 5.0
± 0.2 4.9
± 0.3 4.7
± 0.3 4.8
± 0.3 4.7
± 0.3 4.1 ~ 5.3 Cl (mmol / L) 109.4
± 1.6 110.5
± 1.4 110.2
± 2.2 110.9
± 1.3 110.4
± 2.2 105 ~ 116 * CBC, complete blood count; WBC, white blood cell; RBC, red blood cell; Hb, haemoglobin; HCT, hematocrit; MCV, mean corpuscular volume; MCH, mean corpuacular haemoglobin; MCHC, mean corpuscular haemoglobin concentration; PLT, platelets; TP, total protein; ALB, albumin; BUN, blood urea nitrogen; CREA, creatinine; AST, aspartate transmainase; ALT, alanine aminotransferase; ALP, alkaline phosphatase; Na, sodium; K, potassium; Cl, Chloride.

Urinalysis results, Urine specific gravity (USG), Urine dip stick, Wet preparation and cytology (Cytology) were performed for all test dogs during the experiment period. The urine test results are shown in Table 8 below.

Group 1 (control) Day0 Day7 Day14 Day21 Day28 Urin-alysis S.G. 1.038
± 0.003 1.041
± 0.002 1.042
± 0.001 1.041
± 0.004 1.039
± 0.004 Dip stick - protein trace protein trace - - Wet - - - - - Cytology - - - - - Group 2 (X1) Day0 Day7 Day14 Day21 Day28 Urin-alysis S.G. 1.035
± 0.005 1.038
± 0.004 1.040
± 0.002 1.034
± 0.005 1.036
± 0.005 Dip stick - - protein trace - - Wet - - - - - Cytology - - - - - Group 3 (X4) Day0 Day7 Day14 Day21 Day28 Urin-alysis S.G. 1.045 ± 0.000 1.037 ± 0.006 1.039 ± 0.005 1.040 ± 0.001 1.037 ± 0.004 Dip stick protein trace - - protein trace - Wet - - - - - Cytology - - - - - Group 4 (X16) Day0 Day7 Day14 Day21 Day28 Urin-alysis S.G. 1.042 ± 0.002 1.040 ± 0.001 1.037 ± 0.002 1.036 ± 0.003 1.040 ± 0.001 Dip stick protein trace protein trace - - - Wet - - - - - Cytology - - - - - * S.G., Specific gravity; No remarkable finding was indicated as “-”.

No abnormalities were found on the urine test during the entire experimental period, and protein traces by concentrated urine were confirmed in several individuals, which were evaluated as normal values, and all values were within the reference range.

As a result of the electrocardiogram (Electrocardiography), electrocardiography was performed on all test dogs during the experiment period, and it was confirmed whether arrhythmia, atrial fibrillation, or other abnormal waveforms appeared. In addition, by measuring indicators such as P wave amplitude (mV), duration (s), QRS wave amplitude (mV), QRS duration (s), and T wave amplitude (mV), abnormalities such as heart deformation and bundle branch blocks It was confirmed that it was observed.

The results of the electrocardiogram test are shown in FIGS. 6 to 9. Referring to this result, no abnormality was observed in the electrocardiogram results of the whole experimental dogs during all the experimental periods, and all values were within the reference range.

Echocardiography results were performed before and after the experiment (Day 0) and after the experiment (Day 28), echocardiography was performed on all test dogs to confirm the morphological changes and basic functional indicators of the heart. Presence or absence was checked, and the results are shown in Table 9. EPIQ 7 Cardiology Ultrasound Machine (Philips, USA) was used as the echocardiogram device.

Parameter (units) of Day 0 Dogs (n = 10) Parameter (units) of Day 28 Dogs (n = 10) Ao Vel (m / s) 124.6 ± 31.4 Ao Vel (m / s) 126.3 ± 27.8 Ao * (cm) 1.7 ± 0.1 Ao * (cm) 1.7 ± 0.6 FS (%) 44.1 ± 4.6 FS (%) 51.7 ± 3.5 HR (bpm) 137.5 ± 16.2 HR (bpm) 142.3 ± 16.7 IVSd * (cm) 0.9 ± 0.2 IVSd * (cm) 0.9 ± 0.1 IVSs * (cm) 1.2 ± 0.3 IVSs * (cm) 1.3 ± 0.4 LA * (cm) 2.1 ± 0.1 LA * (cm) 2.0 ± 0.7 LVET (ms) 186.2 ± 8.1 LVET (ms) 198.7 ± 9.2 LVIDd * (cm) 3.1 ± 0.3 LVIDd * (cm) 3.0 ± 0.8 LVIDs * (cm) 1.7 ± 0.4 LVIDs * (cm) 1.6 ± 0.6 LVPWd * (cm) 0.9 ± 0.2 LVPWd * (cm) 0.8 ± 0.2 LVPWs * (cm) 1.3 ± 0.2 LVPWs * (cm) 1.3 ± 0.2 PEP (ms) 21.5 ± 2.8 PEP (ms) 24.8 ± 3.6 MBP (mmHg) 94.7 ± 6.5 MBP (mmHg) 100.5 ± 7.6 DBP (mmHg) 77.5 ± 12.5 DBP (mmHg) 89.0 ± 6.5 SBP (mmHg) 127.8 ± 9.1 SBP (mmHg) 129.5 ± 8.9 Ao Vel, aortic velocity; Ao, aortic diameter; FS, fractional shortening; HR, heart rate; IVSd, interventricular septum (diastole); IVSs, interventricular septum (systole); LA, left atrium; LVET, left ventricular ejection time; LVIDd, left ventricular internal diameter (diastole); LVIDs, left ventricular internal diameter (systole); LVPWd, left ventricular posterior wall (diastole); LVPWs, left ventricular posterior wall (systole); PEP, pre-ejection period; MBP, mean blood pressure; DBP, diastolic blood pressure; SBP, systolic blood pressure.
Values expressed as mean standard ± deviation. * Indexed to body weight (Cornell et al., 2004).

Referring to the results in Table 9, no significant changes in heart size or abnormality were observed for each individual before (0 day) and after (28 day) results of echocardiography results, and all results were normal. Appeared within range.

The necropsy was visually evaluated for organ (cardiac, lung, liver, spleen, kidney, and digestive) toxicities through an autopsy after the 28-day experiment, and the heart was examined for histopathological changes through biopsy. Did. At the time of visual inspection for each organ, the heart, lungs, and both in group 1 (control group), group 2 (recommended dose group), group 3 (4 times over recommended dose group), and group 4 (16 times over recommended dose group) No gross lesions of the liver, spleen, kidneys and digestive system were observed.

No specific lesions were observed in the bicuspid valve and left ventricular outflow tract, and no mucosal degeneration of the bicuspid valve was observed. No specific lesions were observed in the myocardium of the left ventricular papillary muscle or the ventricular septum. The weight of the bicuspid valve was less than 300 mg, and no proliferation of the bicuspid valve or local bleeding at the base of the bicuspid valve were observed. Hyperplasia of the tendonae at the papillary muscle attachment was not observed, and no left ventricular outflow caused endocardial discoloration, left ventricular myocardial necrosis or scarring. In addition, no lesions such as hematomas of the tricuspid valve were observed. Group 4 (16-fold overdose group recommended dose) The results of a long-term visual examination through necropsy of the experimental dog were attached to FIG. 10 with pictures of the heart and kidneys.

In addition, it was confirmed through histological examination of all experimental dogs that there was a histopathological change, and the results are shown in FIG. 11. Group 1 (control group), group 2 (recommended dose group), group 3 (recommended 4-fold overdose group), and group 4 (recommended 16-fold overdose group) mucus-species degeneration and proliferation of bicuspid valves, left ventricular outflow No proliferation of the endocardium, multiple necrosis of the left ventricle and ventricular septal myocardium, and proliferation of the arterial wall in the left ventricle were not observed.

In conclusion, the composition of Example 2 above did not show specific gross organ abnormalities and myocardial histopathological abnormalities not only in the recommended dose but also in the 4 and 16-fold overdose groups.

When synthesizing the above test results, the composition of Example 2 of the present invention, the recommended dose (0.25 mg per body weight based on pimobendan), 4 times the recommended dose (1 mg twice per body weight based on pimobendan) ), 16 times the recommended dose (4 mg twice per day based on pimobendan) with various toxic doses, when administered for a total of 4 weeks, the dog does not show any specific clinical symptoms including dysfunction, hypersensitivity, digestive symptoms, etc. No specific side effects or evidence of toxicity were found on basic physical examination, blood test, urine test, electrocardiogram test, echocardiographic results, visual examination through autopsy, and biopsy. Therefore, the above composition is considered a safe drug that can be used without specific side effects when administered to dogs.

4. Short-term clinical evaluation

(1) Experiment method

In dogs with naturally occurring bicuspid valve degeneration (MMVD), the safety and effectiveness of the composition of the present invention was assessed on a small scale in a short period of time compared to conventional drugs.

A shaft axis with myxomatous mitral valve degeneration was recruited and participated. The experiment included one control group to which the conventional drug Pimobendan (Vetmedin®, Boeringer Ingelheim, Germany) was administered, and one experimental group to administer the prototype of Example 2 prepared in the present invention.

Inclusion criteria for the control group and the experimental group are dogs over 1 year of age diagnosed with Myxomatous mitral valve degeneration in the veterinary clinic. Blood reflux should be checked. In addition, among the individuals with Stage B or higher classification of the American Veterinary Internal Medicine Association (ACVIM) chronic valve heart disease, it was limited to those instructed to administer Pimobendan. Exclusion criteria are dogs with other heart disease (eg, congenital heart disease) other than bicuspid valve degeneration (MMVD), and individuals with severe systemic diseases other than heart disease (physical examination, blood and serochemistry) have also been excluded. In addition, congestion prescribed for medications other than heart-related medications was excluded.

Individuals selected for small-scale experiments (n = 2) were randomly divided into a control group (existing drug-administered group) and an experimental group (composition-administered group of Example 2), and the experiment was conducted and evaluated by the following items.

-Physical examination (dehydration, physical fitness index, weight, body temperature, heart rate, respiratory rate, blood pressure)

-Blood test (whole blood cell test: CBC (WBC, HCT, Plt, serochemical test: ALT, AST, ALP, TP, Alb, BUN, Cre, electrolyte test: Na, K, Cl)

-Image evaluation (radiation evaluation, electrocardiogram evaluation, echocardiography evaluation)

-Echocardiography (MV prolapse (Y / N), FS, EF, LVIDd, LVIDs, ESVI, EDVI, LA / Ao, AT (RVOTflow), AT / RVET, E, A, E / A, Em, Am, E / Em], Efficacy Assessment (Heart Index Scorecard and Efficacy Scorecard and Score Assessment)]

* Based on the scorecard

-Electrocardiogram (no abnormality, arrhythmia observation, atrial fibrillation observation, other abnormal waveform observation)

-Radiographic examination (heart size, VHS; no cardiac hypertrophy, weak cardiac hypertrophy, moderate cardiac hypertrophy, severe cardiac hypertrophy),

-Evaluation of wastewater species (no wastewater species, weak increase in lung parenchyma, moderate increase in lung parenchyma, presence of Alveolar pattern, presence of severe consolidation)

-Clinical efficacy after drug administration (clinical symptoms improved significantly, clinical symptoms improved, clinical symptoms slightly improved, clinical symptoms did not improve, treatment failed),

-Side effects (with or without).

* Based on the effectiveness evaluation table

-Appetite: 1 increase, 2 normal, 3 decrease (2/3 of normal), 4 significant decrease (less than 2/3 of normal)

-Behavior: 1 sensitive and responsive, 2 slightly depressed, 3 severely depressed, 4 minimally responsive, 5 unresponsive

-Exercise intolerance: 1 very good, 2 good (slightly reduced running ability), 3 reduced (inactive than normal, trying to avoid long workouts), 4 poor (almost no movement other than eating, drinking, urination)

-Respiration status evaluation: 1 normal, 2 slightly increased breathing effort, 3 severity of breathing difficulty, 4 severe breathing difficulty)

-Cough: 1 None, 2 Sometimes (several times a week), 3 Frequently (several times a day), 4 sustained (several times a day)

-Night breathing symptoms: 1 None, 2 Sometimes coughing at night, but no breathing difficulties, 3 Coughing at night, uncomfortable breathing

-Heart failure stage, ISACHC: 1 Symptoms of heart disease but no symptoms of volumetric or pressure overload ventricular hypertrophy; 2 Symptoms of heart disease are visible and compensation can be detected by echocardiography or radiation; 3 At rest or during light exercise, symptoms of heart failure are evident, deteriorating quality of life. Typical symptoms of heart failure include exercise intolerance, coughing, frequent breathing, mild respiratory distress, and mild to moderate revenge. Hypoperfusion at rest does not usually appear; 4 Symptoms of advanced congestive heart failure are obvious. Clinical symptoms include dyspnea, pronounced ascites, severe exercise intolerance, or hypoperfusion at rest. Hospital treatment (home treatment) available; 5 Inpatient treatment is essential (psychic shock, life-threatening pulmonary edema, massive pleural effusion).

-Side effects evaluation

Systemic reactions, skin reactions, digestive symptoms, presence of hematologic abnormalities, presence of serological abnormalities, abnormalities in urine tests, cardiac drug relevance, first aid and treatment, evaluation of recovery items

Subjects were evaluated prior to dosing (day 0) and 7 days post dosing starting from the start date of the experiment.

(2) Experimental results

-Basic information and physical / blood test results

Table 10 shows the basic information and physical examination results of the axial axis included in the control group and the experimental group.

Information Control Experimental group Breed Pekignese Schunauzer Age (years) 10 8 Gender Intact female Intact female History of present illness Heart noise present without clinical symptoms Heart noise present without clinical symptoms Item Control Experimental group General condition Mild depression NRF Body condition score 3 out of 5 3 out of 5 Body weight (Kg) 6.62 7.6 Body temperature (℃) 38.8 38.7 Heart rate (bpm) 138 126 Respiratory rate (/ min) 18 36 Blood pressure S 146 D 89 MAP 120 S 142 D 95 MAP 117 * NRF, no remarkable findings; S, systolic blood pressure; D, diastolic blood pressure; MAP, mean arterial pressure.

In the control group and the experimental group, the blood test (whole blood cell test, serochemical test, electrolyte test) was performed on day 0 and day 7 of the experiment, and the results are shown in Table 11 below.

Evaluation items Control Experimental group Reference range Day 0 Day 7 Day 0 Day 7 CBC WBC (x10 ³ / ul) 13.61 11.15 13.02 12.85 5.2 ~ 13.9 RBC (x10 ⁶ / ul) 5.78 6.37 7.94 8.35 5.7 ~ 8.8 Hb (gm / dl) 13.6 15.7 17.6 18.2 12.9 ~ 18.4 HCT (%) 44.3 46.2 49.7 51.2 37.1 ~ 57.0 MCV (fL) 64.5 72.5 62.6 62.3 58.8 ~ 71.2 MCH (pg) 23.5 24.6 22.2 21.8 20.5 ~ 24.2 MCHC (%) 36.5 34 35.4 35.5 31.0 ~ 36.2 PLT (x10 ³ / ul) 486 411 432 476 143 ~ 400 Serum
Chemistry TP (g / dL) 7.3 7.1 7.2 7.1 5.4 ~ 7.4 ALB (g / dL) 3.4 3.4 3.2 3.3 2.9 ~ 4.2 BUN (mg / dL) 18 13 19 23 8 ~ 26 CREA (mg / dL) 0.8 0.6 One 0.9 0.5 ~ 1.3 AST (U / L) 32 43 34 36 15 ~ 43 ALT (U / L) 56 61 116 87 19 ~ 70 ALP (U / L) 66 52 129 116 15-127 Na (mmol / L) 146 145 153 153 144 ~ 154 K (mmol / L) 4.3 4.3 4.4 4 4.1 ~ 5.3 Cl (mmol / L) 116 115 111 109 105 ~ 116 * CBC, complete blood count; WBC, white blood cell; RBC, red blood cell; Hb, haemoglobin; HCT, hematocrit; MCV, mean corpuscular volume; MCH, mean corpuacular haemoglobin; MCHC, mean corpuscular haemoglobin concentration; PLT, platelets; TP, total protein; ALB, albumin; BUN, blood urea nitrogen; CREA, creatinine; AST, aspartate transmainase; ALT, alanine aminotransferase; ALP, alkaline phosphatase; Na, sodium; K, potassium; Cl, Chloride.

Referring to Table 11 above, in the control group and in the experimental group, blood test results performed on day 0 before administration and day 7 after administration were within the reference range in all values.

-Video evaluation results

Chest radiographic evaluation and echocardiography were performed, and the results are shown in FIGS. 12 to 14.

* Control results

-Degeneration of the bicuspid valve was confirmed.

-Reversal of bicuspid plate on Color Doppler was confirmed

-At day 0, left atrial dilatation (LA / Ao = 1.64) was confirmed, and on day 7 after drug administration, LA / Ao = 1.54.

-The left ventricular shortening rate (FS) was increased to 43.8% on day 0, and increased to 48.1% on day 7, and the ejection rate (EF) was also increased from 76.9% on day 0 to 81.3% on day 7.

-Left ventricular diastolic diameter (LVIDd) decreased slightly from 2.65 cm on Day 0 to 2.58 cm on Day 7 and left ventricular systolic diameters (LVIDs) also decreased from 1.49 cm on Day 0 to 1.34 cm on Day 7.

-The end-diastolic volume index (ESVI) decreased from 5.95 on day 0 to 4.5 on day 7, and the end-diastolic volume index (EDVI) slightly decreased from 25.8 on day 0 to 24.1 on day 7.

-Accelerated time (AT) of pulmonary arterial systolic blood was 87 ms on day 0 and 88 ms on day 7, the likelihood of pulmonary arterial hypertension was low. All of them were 0.4, and the possibility of pulmonary arterial hypertension was also low.

-The E / A ratio of Trans-mitral flow was 1.2, and the Em / Am of Tissue-Doppler imaging was 0.63, and the diastolic dysfunction could be suspected. This was similar on the 7th day.

-Other echocardiography showed no abnormalities or unusual features / side effects of the heart structure.

* Results of the experimental group

-Degeneration of the bicuspid valve was confirmed.

-Reversal of bicuspid plate on Color Doppler was confirmed

-At day 0, left atrial dilatation (LA / Ao = 1.67) was confirmed, and at day 7 after drug administration, LA / Ao = 1.57.

-The rate of left ventricular shortening (FS) was 49.23% on day 0, and increased to 55.4% on day 7, and the ejection rate (EF) also increased from 82.8% on day 0 to 87.3% on day 7.

-Left ventricular diastolic diameter (LVIDd) decreased slightly from 3.23 cm on day 0 to 2.85 cm on the 7th day examination, and left ventricular systolic diameter (LVIDs) also decreased from 1.64 cm on day 0 to 1.27 cm on the 7th day examination.

-The end-diastolic volume index (ESVI) decreased from 4.87 on day 0 to 3.91 on day 7, and the end-diastolic volume index (EDVI) slightly decreased from 34.2 on day 0 to 30.9 on day 7.

-Accelerated time (AT) of pulmonary arterial systolic blood was 87 ms on day 0 and 89 ms on day 7, the probability of pulmonary arterial hypertension was low. All of them were 0.4, and the possibility of pulmonary arterial hypertension was also low.

-The E / A ratio of trans-mitral flow was 1.34, and the Em / Am of Tissue-Doppler imaging was 0.50, and the diastolic dysfunction could be suspected. This was similar on the 7th day.

-Other echocardiography showed no abnormalities or unusual features / side effects of the heart structure.

The results of echocardiography between the control group and the experimental group are shown in Table 12 below.

Variables Control Experimental group Day 0 Day 7 Day 0 Day 7 Echocardio-graphy MV prolapse (Y / N) N N N N FS (%) 43.8 48.1 49.23 55.4 EF (%) 76.9 81.3 82.8 87.3 LVIDd (cm) 2.65 2.58 3.23 2.85 LVIDs (cm) 1.49 1.34 1.64 1.27 ESVI 5.95 4.5 4.87 3.91 EDVI 25.8 24.1 34.2 30.9 LA / Ao 1.64 1.54 1.67 1.57 AT (RVOT flow) (ms) 87 88 87 89 AT / RVET 0.4 0.4 0.4 0.4 E (cm / s) 68 81 58 75 A (cm / s) 56 68 43 58 E / A 1.2 1.2 1.34 1.29 Em (cm / s) 4.58 7.86 5.95 7.45 Am (cm / s) 7.26 10.6 8.75 14.7 E / Em 14.8 10.3 9.88 10.12 * MV, mitral valve; FS, fractional shortening; EF, ejectional fraction; LVIDd, left ventricular internal diameter (diastole); LVIDs, left ventricular internal diameter (systole); ESVI, end systolic volume index; EDVI, end diastolic volume index; LA, left atrium; Ao, Aorta; AT, acceleration time; RVOT, right ventricular outflow tract; RVET, right ventricular ejection time; E, e peak flow velocity of transmitral flow; A, a peak flow velocity of transmitral flow; Em, e peak flow velocity of tissue doppler imaging; Am, a peak flow velocity of tissue doppler imaging.

Summarizing the above results, both the control group and the experimental group showed a similar trend in the same index after drug administration in both the constriction, and confirmed an increase in contractility and improvement in heart size.

-ECG test results

Electrocardiography was performed on day 0 before drug administration and on day 7 after administration, and the results are shown in FIGS. 15 and 16.

15 and 16, the ECG test performed in the control axis showed normal sinus rhythm, and the QRS amplitude was 2.8 mV, which showed a slightly increased QRS amplitude when considering a small dog, which would suggest left ventricular expansion. Can be. All other values were within the normal range. As a result of the ECG test performed in the experimental group, the normal sinus rhythm was shown, and the QRS amplitude was 4 mV, showing a slightly increased QRS amplitude, considering small size, which may suspect left ventricular expansion. All other values were within the normal range.

No significant ECG changes or abnormalities were observed before and after drug administration in both control and experimental groups.

-Evaluation of effectiveness

The items and scores of the efficacy evaluation table were evaluated for each patient in the control group and the experimental group, and the results are shown in Table 13 below.

Referring to Table 13 above, the sum of the scores on the efficacy evaluation table in the control axis was 9 points on Day 0, and was also 9 points on Day 7 after administration. This is considered to be difficult to clearly observe the improvement of the clinical symptoms since this patient is an ACVIM B2 patient without clinical symptoms. The sum of the efficacy scores in the experimental group reduction was 9 points on Day 0 and decreased to 8 points on Day 7 after dosing. It is believed that the ectopic cough that was present intermittently improved after administration of the composition of the present invention.

-Heart Indicator Scorecard

To check whether the cardiac index was improved before and after drug administration, the cardiac index of the control group and the experimental group was scored according to the following Table 14 items.

score electrocardiogram Radiography
(Heart size, VHS) Radiography
(Evaluation of waste species) Clinical efficacy after drug administration Side effects One nothing strange No cardiac hypertrophy No waste species Clinical symptoms improved significantly No side effects 2 Arrhythmia observation Weak cardiomegaly Lung parenchyma slightly increased Clinical symptoms improved Side effects exist 3 Atrial fibrillation observation Moderate Symbiosis Moderate increase in lung parenchyma Clinical symptoms improved slightly - 4 Other abnormal waveform observation Severe cardiomegaly Alveolar pattern present Clinical symptoms do not improve - 5 - - Severe consolidation Treatment failure -

The score of each patient according to the scorecard was scored in the heart index of the control group: 9 points; The cardiac index score of the experimental group reduction was 9 points, which was the same.

-Side effects evaluation

In the control group and the experimental group, the side effects were judged according to the side effect evaluation table, and no side effects were indicated in all items.

(3) Evaluation of efficacy and safety between drugs

-Effectiveness evaluation method

The criteria for evaluating the effectiveness of the composition of Example 2 administered to the control group and the existing drug administered to the control group are as follows.

1) Improvement rate of effectiveness evaluation score

2) Heart score

3) Heart size improvement rate (LA / Ao, left ventricular diastolic diameter)

4) Left ventricular shortening rate improvement rate

-Results of effectiveness evaluation

The effectiveness evaluation score and the improvement rate of the heart size index are calculated as follows.

Improvement rate (%) = (day 0 result-day 7 result) ÷ (day 0 result) x 100 (%)

(However, in the case of left ventricular contraction rate, the position of the 7th and 0th results is changed.)

In Table 15, the effectiveness of the control group and the experimental group was compared and evaluated.

Evaluation item Control Experimental group Effectiveness score improvement rate (%) 0 11 Heart Indicator Score (Point) 9 9 Heart size improvement rate (%) LA / Ao improvement rate 6 6 Left ventricular diastolic diameter improvement rate 3 12 Left ventricular shortening rate (FS) improvement rate 9 11

In total, the subjects who participated in the test were divided into two groups, a control group (existing drug administration group) and an experimental group (composition of the present invention). The control group was Pekignese, a 10-year-old intact female, and had no heart murmur without clinical symptoms. The experimental group was Schunauzer, an 8-year-old intact female, who also had noisy symptoms without clinical symptoms.

Basic physical examinations such as dehydration, body loss index, body weight, body temperature, heart rate, respiratory rate, and blood pressure were performed on both axes. And all other items were within the normal range. The experimental group also showed hypertensive blood pressure with systolic blood pressure of 142 mmHg, and all other items were within the normal range. This is likely due to a compensatory increase in blood pressure due to heart disease in both axes. Blood test (whole blood cell test, serum chemistry test, electrolyte test) was performed on the 0th and 7th day of experiment in the control group and the experimental group. Results were within the reference range for all figures. Therefore, 0 ~ 10% or less of side effects compared to the existing drug (pimobendan), which was the quantitative goal of this experiment-Evaluation items: Blood test (blood, serum chemistry test, especially liver and kidney level, electrolyte level evaluation), frequency of occurrence of side effects, and kind evaluation- Was satisfied.

The results of chest radiographic evaluation performed in the control group showed a weak bronchial-epileptic pattern throughout the lung field, and left and right heart hypertrophy could be suspected. The results of chest radiographic evaluation conducted in the experimental group also showed a weak or severe bronchial-epileptic pattern throughout the lung field, and no other specific findings were observed in both axes. Echocardiography was performed on the 0th and 7th days in the control group and the experimental group, and both circumferences showed a similar trend in the same index after administration of the drug (conventional drug and composition of the present invention), increased contractility, and improved heart size Was confirmed.

Electrocardiography was performed on day 0 before drug administration and on day 7 after administration, and the ECG test performed in the control axis showed normal sinus rhythm, and the QRS amplitude was 2.8 mV. And this may suspect the expansion of the left ventricle. All other values were within the normal range. As a result of the ECG test performed in the experimental group, the normal sinus rhythm was shown, and the QRS amplitude was 4 mV, showing a slightly increased QRS amplitude, considering small size, which may suspect left ventricular expansion. All other values were within the normal range. No significant ECG changes or abnormalities were observed before and after drug administration in both control and experimental groups.

In the control group, the sum of the scores on the EFF was 9 points on Day 0, and was also 9 points on Day 7 after administration. This is considered to be difficult to clearly observe the improvement of the clinical symptoms since this patient is an ACVIM B2 patient without clinical symptoms. The sum of the efficacy scores in the experimental group reduction was 9 points on Day 0 and decreased to 8 points on Day 7 after dosing. It is believed that the ectopic cough that was present intermittently improved after the composition administration of the present invention. In order to confirm whether the cardiac index was improved before and after drug administration, the cardiac index of the control group and the experimental group was scored, and each patient's score according to the scorecard was the same at 9 points. In the control group and the experimental group, the side effects were judged according to the side effect evaluation table, and no side effects were indicated in all items.

For evaluating the effectiveness of the existing drug administered to the control group and the composition drug of Example 2 administered to the experimental group 1) efficacy evaluation score improvement rate, 2) heart index score, 3) heart size improvement rate (LA / Ao, left ventricular diastolic diameter), 4) The rate of improvement of the left ventricular shortening rate was evaluated. As a result of the efficacy evaluation, the improvement rate of the efficacy evaluation score was 0% in the control group and 11% in the experimental group, and the experimental group showed a significant improvement rate. In the case of the heart size improvement rate, the LA / Ao improvement rate was 6% in the control group and 6% in the experimental group, ie, the improvement rate of the composition of the present invention was 0% compared to the existing drug. In the case of the left ventricular diastolic diameter improvement rate, 3% in the control group and 12% in the experimental group showed a significant improvement rate compared to the conventional drug. In addition, the improvement rate of the left ventricular shortening rate (FS) was 9% in the control group and 11% in the experimental group, showing a significant improvement rate of 22% compared to the existing drug. This is a quantitative goal of this experiment, the evaluation of efficacy-Improvement of heart disease 0 ~ 10% compared to the existing drug (pimobendan)-Evaluation items: clinical symptom scorecard, cardiac index scorecard and heart size (LA / Ao ratio, left ventricular diastolic diameter)- Was satisfied.

Synthesizing the above results, it was confirmed that the pimobendan and pentoxifylline mixture composition developed in this experiment has an effect equal to or higher than that of the existing drug, and has a positive effect of improving heart function in actual heart disease reduction. In addition, no side effects were observed in clinical symptoms such as clinical symptoms, physical examination, radiation, echocardiography, electrocardiogram, etc., and thus it was confirmed that it has an equivalent or higher safety compared to existing drugs.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also provided. It belongs to the scope of rights.

Claims (11)

Pimobendan or a pharmaceutically acceptable salt thereof;
Pentoxifylline or a pharmaceutically acceptable salt thereof; And
A palatable excipient selected from the group consisting of beef powder, chicken liver powder, and combinations thereof; containing, oral administration preparations for the treatment or symptom relief of canine valve degeneration. According to claim 1,
The pimobendan or a pharmaceutically acceptable salt thereof is contained in 0.005 to 0.02 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof, to treat or treat symptoms of bicuspid valve degeneration in canine animals. For oral administration formulations. According to claim 1,
The oral administration formulation further comprises a sweetener, oral administration formulations for the treatment or symptom relief of bicuspid valve degeneration in canine animals. According to claim 1,
The oral administration formulation further comprises an anti-caking agent, an oral administration formulation for the treatment or symptom relief of bicuspid valve degeneration in canine animals. According to claim 1,
The palatable excipients are contained in 1 to 20 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof, orally administered preparations for the treatment or symptom relief of canine valve degeneration. According to claim 3,
The sweetener is contained in 0.1 to 3 parts by weight based on 1 part by weight of the pentoxifylline or a pharmaceutically acceptable salt thereof, orally administered formulation for the treatment or symptom relief of bicuspid valve degeneration in canine animals. According to claim 1,
The oral dosage form is a powder, which is administered twice a day by dividing 0.3 to 0.8 g / kgBW / day, orally administered preparation for treating or reducing symptoms of bicuspid valve degeneration in canine animals. According to claim 1,
The oral administration formulation, which improves peripheral blood circulation, has a blood circulation improving activity including an increase in coronary artery blood flow in the heart, orally administered formulation for the treatment or symptom relief of canine valve degeneration in canine animals. According to claim 1,
The oral dosage form is an improved rate of improvement of the diameter of the left ventricular diastolic in a canine animal with bicuspid valve degeneration, an oral dosage form for treating or reducing symptoms of bicuspid valve degeneration in a canine animal. According to claim 1,
The oral dosage formulation is to reduce the degree of coughing in canine animals with bicuspid valve degeneration, orally administered formulations for the treatment or symptom relief of bicuspid valve degeneration in canine animals. According to claim 1,
The oral dosage form is to be added to the feed of the canine animal feed, oral canal preparations for the treatment or symptom relief of canine valve degeneration.

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