KR20200083094A - Cyproheptadine derivatives, process for preparing the same and composition for promoting appetite comprising the same - Google Patents

Abstract

The present invention provides a cyproheptadine derivative or a pharmaceutically acceptable salt thereof, a method for manufacturing the same, and a composition for promoting appetite of mammals containing the same. The cyproheptadine derivative of the present invention exhibits a similar or lower AgRP expression level to cyproheptadine in a mHypoE cell line to exhibit an excellent appetite promoting effect, and also exhibits an excellent gaining effect in broilers and piglets. Therefore, the cyproheptadine derivative or the pharmaceutically acceptable salt thereof, the method for manufacturing the same, and the composition for promoting appetite of mammals can be usefully used in the field of animal medicines and feed additives.

Description

Ciproheptadine derivatives, method for manufacturing same, and composition for promoting appetite comprising the same{Cyproheptadine derivatives, process for preparing the same and composition for promoting appetite comprising the same}

The present invention relates to a ciproheptadine derivative and a method for preparing the same, and more specifically, a novel ciproheptadine derivative or a pharmaceutically acceptable salt thereof, a method for preparing the same, and a composition for promoting appetite in mammals comprising the same It is about.

In the animal industry, loss of appetite leads to an increased probability of secondary infections due to reduced productivity and immunity. Therefore, a method for promoting appetite of industrial animals has been developed to increase productivity as well as increase immunity and prevent infection.

Most of the appetite accelerators of industrial animals commercially available in Korea are injections, and there is inconvenience in the route of administration. Therefore, it is necessary to develop oral feed additives through the appetite central nervous system.

Existing feed additive products employ a mechanism that induces an indirect effect by adding a flavoring agent, a sweetener, or the like, but these products have a problem of insufficient appetite improvement effect.

US Patent Publication US20020019533A1 (2002.02.14.) International patent WO198912443A1 (1989.12.28.) United States Patent US5095022A1 (1992.03.10.) International patent WO200024715A1 (2000.05.04.)

The inventors of the present invention, while studying for the development of appetite-enhancing animal medicines, focused on the fact that drugs with high histamine H1 receptor affinity among psychotic drugs exhibit a remarkable weight gain effect, histamine through molecular modeling activity prediction By designing cyproheptadine derivatives with H1 receptor affinity and evaluating their appetite promoting effects through molecular modeling and bioactivity experiments and animal experiments, ciproheptadine derivatives of the novel structure according to the present invention It has been confirmed that can be used for the purpose of promoting appetite in mammals.

Accordingly, an object of the present invention is to provide a novel ciprofheptadin derivative or a pharmaceutically acceptable salt thereof, a method for preparing the same, and a composition for promoting appetite in mammals comprising the same.

According to an aspect of the present invention, there is provided a compound of Formula 1, or a pharmaceutically acceptable salt thereof.

<Formula 1>

In the above formula, X is R ₁ , R ₂ , COR ₃ , or COR ₄ ,

R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ to C ₉ alkyl, substituted or unsubstituted C ₃ to C ₉ alkenyl, substituted or unsubstituted C ₃ to C ₉ alkynyl, or Dioxolanylalkyl,

R ₂ is alkoxybenzyl or alkylpyridine,

R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, phenylketonyl, or phenylalkynyl,

R ₄ is substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl.

In one embodiment, R ₁ may be benzyl, phenylpropyl, cinnamil, nonyl, allyl, butenyl, methylbutenyl, propynyl, or dioxolanylmethyl, and R ₂ is methoxybenzyl or methylpyridine May be, the R ₃ may be cyclopentyl, cyclohexenyl, ethylacetyl, dihydroxyphenyl, methylphenylketonyl, or phenylethynyl, and R ₄ may be hexyl, tolyl, or benzyl.

Specific examples of preferred compounds as compounds according to the invention are as follows:

(1) 1-Benzyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine

(2) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-phenylpropyl)piperidine

(3) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(4-methoxybenzyl)piperidine]

(4) 1-cinnamyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine

(5) 3-((4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)methyl)pyridine

(6) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-nonylpiperidine

(7) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(prop-2-ain-1-yl)piperidine

(8) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-methylbut-2-en-1-yl)piperidine

(9) 1-allyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine

(10) (E) -1- (boot-2-en-1-yl) -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine

11 1 - ((1,3-dioxolan-2-yl) methyl) -4- (5 H-dibenzo [a, d] [7] annulene-5-ylidene) piperidine

(12) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (cyclopentyl) methanone

(13) 1- (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) octane-1-one

(14) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (cyclo hex-1-en-1-yl) -methanone

(15) Ethyl 3-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-oxopropanoate

(16) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (3,5-di-hydroxyphenyl) methanone

(17) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-2-phenylethane-1,2-dione

(18) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (p- tolyl) methanone

(19) 1- (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl carbonyl) -2-phenyl-ethane-1-one

(20) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-phenylprop-2-ain-1 -On

According to another aspect of the present invention, (a) by removing a methyl group from cyproheptadine to synthesize 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine step; And (b) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and R ₁ -Br, R ₂ -Cl, R ₃ -COOH and R _4- Reacting one selected from the group consisting of COCl (R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ to C ₉ alkyl, substituted or unsubstituted C ₃ to C ₉ eggs) Kenyl, substituted or unsubstituted C ₃ to C ₉ alkynyl, or dioxolanylalkyl; R ₂ is alkoxybenzyl or alkylpyridine; R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, Phenylketonyl, or phenylalkynyl; R ₄ is substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl) is provided.

In one embodiment, in step (b), 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine can be reacted with R ₁ -Br, or 4 -(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine can be reacted with R ₂ -Cl, or 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine can be reacted with R ₃ -COOH, or 4-(5 H -dibenzo[ a,d ][7]anulen-5-yl Den) piperidine and R ₄ -COCl can be reacted.

According to another aspect of the present invention, there is provided a composition for promoting appetite in mammals comprising the compound as an active ingredient.

In one embodiment, the composition for promoting appetite may be an animal medicine or feed additive.

It has been found that the novel ciproheptadine derivatives designed in the present invention show agRP expression similar to or lower than that of ciproheptadine in the mHypoE cell line, thereby showing an excellent appetite promoting effect, and an excellent weight gain improving effect in broilers and piglets.

Therefore, the compound according to the present invention, i.e., a ciproheptadine derivative or a pharmaceutically acceptable salt thereof, a method for preparing the same, and a composition for promoting appetite in mammals comprising the same can be usefully used in the field of animal medicine and feed additives .

1 is a ¹ H NMR spectrum of CYP001 (Example 1).
2 is a ¹ H NMR spectrum of CYP002 (Example 2).
3 is a ¹ H NMR spectrum of CYP003 (Example 3).
4 is a ¹ H NMR spectrum of CYP004 (Example 4).
5 is a ¹ H NMR spectrum of CYP005 (Example 5).
6 is a ¹ H NMR spectrum of CYP006 (Example 6).
7 is a ¹ H NMR spectrum of CYP007 (Example 7).
8 is a ¹ H NMR spectrum of CYP008 (Example 8).
9 is a ¹ H NMR spectrum of CYP009 (Example 9).
10 is a ¹ H NMR spectrum of CYP010 (Example 10).
11 is a ¹ H NMR spectrum of CYP011 (Example 11).
12 is a ¹ H NMR spectrum of CYP013 (Example 12).
13 is a ¹ H NMR spectrum of CYP014 (Example 13).
14 is a ¹ H NMR spectrum of CYP015 (Example 14).
15 is a ¹ H NMR spectrum of CYP016 (Example 15).
16 is a ¹ H NMR spectrum of CYP018 (Example 16).
17 is a ¹ H NMR spectrum of CYP019 (Example 17).
18 is a ¹ H NMR spectrum of CYP020 (Example 18).
19 is a ¹ H NMR spectrum of CYP021 (Example 19).
20 is a ¹ H NMR spectrum of CYP022 (Example 20).
21 is a result showing the effect of reducing the amount of expression of AgRP (Agouti-related peptide) of ciproheptadine.
22 is a result showing the effect of reducing the amount of AgRP expression of ciproheptadine derivatives.
23 is a result showing the effect of reducing the amount of AgRP expression of the selected five candidate substances (CYP002, CYP006, CYP008, CYP014, CYP022).
Fig. 24 shows the results of the analysis of cyproheptadine with established HPLC conditions.
25 is a result of analyzing CYP002 under established HPLC conditions.
26 shows the results of analyzing CYP006 under established HPLC conditions.
27 is a result of analyzing CYP022 with established HPLC conditions.
Fig. 28 is a photograph of a new compound feed additive performing broiler weight increase experiments (Cyproheptadine(a), CYP 002(b), CYP 006(c), CYP 022(d)).
29 is a photograph of the configuration of a test group that performed a broiler weight gain experiment.
Fig. 30 is the average weight gain curve of each group broiler fed the feed supplemented with each test substance [*p<0.05, significantly different from the negative control].
Fig. 31 is the average weekly weight change of broilers in each group fed with the feed added with each test substance [*p<0.05, significant difference from negative control].
Figure 32 is serum cortisol concentration at 28 days after treatment of each test substance with feed additives.
33 is a photograph showing the configuration of the test group, (a) is a negative administration group, (b) is a CYP022 administration group (left) and a positive control (right).

The present invention provides a compound of Formula 1, or a pharmaceutically acceptable salt thereof.

<Formula 1>

In the above formula, X is R ₁ , R ₂ , COR ₃ , or COR ₄ ,

R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ to C ₉ alkyl, substituted or unsubstituted C ₃ to C ₉ alkenyl, substituted or unsubstituted C ₃ to C ₉ alkynyl, or Dioxolanylalkyl,

R ₂ is alkoxybenzyl or alkylpyridine,

R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, phenylketonyl, or phenylalkynyl,

R ₄ is substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl.

In one embodiment, R ₁ may be benzyl, phenylpropyl, cinnamil, nonyl, allyl, butenyl, methylbutenyl, propynyl, or dioxolanylmethyl, and R ₂ is methoxybenzyl or methylpyridine May be, the R ₃ may be cyclopentyl, cyclohexenyl, ethylacetyl, dihydroxyphenyl, methylphenylketonyl, or phenylethynyl, and R ₄ may be hexyl, tolyl, or benzyl.

Specific examples of preferred compounds as compounds according to the invention are as follows:

(1) 1-Benzyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine [1-Benzyl-4- (5 H -dibenzo [a, d ][7]annulen-5-ylidene)piperidine]

(2) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-phenylpropyl)piperidine [4-(5 H -Dibenzo[ a, d ][7]annulen-5-ylidene)-1-(3-phenylpropyl)piperidine]

(3) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(4-methoxybenzyl)piperidine [4-(5 H -Dibenzo[ a ,d ][7]annulen-5-ylidene)-1-(4-methoxybenzyl)piperidine]

(4) 1-cinnamyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine [1-Cinnamyl-4- (5 H -dibenzo [a, d ][7]annulen-5-ylidene)piperidine]

(5) 3-((4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)methyl)pyridine [3-((4-( 5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)methyl)pyridine]

(6) 4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) -1-piperidine furnace nilpi [4- (5 H -Dibenzo [a , d] [7] annulen-5-ylidene)-1-nonylpiperidine]

(7) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(prop-2-ain-1-yl)piperidine [4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)-1-(prop-2-yn-1-yl)piperidine]

(8) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-methylbut-2-en-1-yl)piperidine [4- (5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)-1-(3-methylbut-2-en-1-yl)piperidine]

(9) 1-allyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine [1-Allyl-4- (5 H -dibenzo [a, d ][7]annulen-5-ylidene)piperidine]

(10) (E) -1- (boot-2-en-1-yl) -4-piperidine (5 H dibenzo [a, d] [7] annulene-5-ylidene) - ( E )-1-(But-2-en-1-yl)-4-(5 H -dibenzo[ a,d ][7]annulen-5-ylidene)piperidine]

(11) 1-((1,3-dioxolan-2-yl)methyl)-4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine [1 -((1,3-Dioxolan-2-yl)methyl)-4-(5 H -dibenzo[ a,d ][7]annulen-5-ylidene)piperidine]

(12) (4-(5 H -dibenzo[a,d][7]anulen-5-ylidene)piperidin-1-yl)(cyclopentyl)methanone [(4-(5 H- Dibenzo[a,d][7]annulen-5-ylidene)piperidin-1-yl)(cyclopentyl)methanone]

(13) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)octan-1-one [1-(4-( 5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)octan-1-one]

(14) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (cyclo hex-1-en-1-yl) -methanone [(4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)(cyclohex-1-en-1-yl)methanone]

(15) Ethyl 3-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-oxopropanoate [Ethyl 3- (4-(5 H -dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)-3-oxopropanoate]

(16) (4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)(3,5-dihydroxyphenyl)methanone [( 4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)(3,5-dihydroxyphenyl)methanone]

(17) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-2-phenylethane-1,2-dione [ 1-(4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)-2-phenylethane-1,2-dione]

(18) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (p- tolyl) methanone [(4- (5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)(p-tolyl)methanone]

(19) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-2-phenylethan-1-one [1- (4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)-2-phenylethan-1-one]

(20) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-phenylprop-2-ain-1 -One [1-(4-(5 H -Dibenzo[ a,d ][7]annulen-5-ylidene)piperidin-1-yl)-3-phenylprop-2-yn-1-one]

The present invention further comprises: (a) synthesizing 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine by removing a methyl group from cyproheptadine; And (b) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and R ₁ -Br, R ₂ -Cl, R ₃ -COOH and R _4- Reacting one selected from the group consisting of COCl (R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ to C ₉ alkyl, substituted or unsubstituted C ₃ to C ₉ eggs) Kenyl, substituted or unsubstituted C ₃ to C ₉ alkynyl, or dioxolanylalkyl; R ₂ is alkoxybenzyl or alkylpyridine; R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, Phenylketonyl, or phenylalkynyl; R ₄ is a substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl).

In the production method of the present invention, step (a) synthesizes 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine by removing methyl groups from ciprofeptahdine. It is a step. After adjusting the pH of the ciproheptadine salt compound to obtain free ciproheptadine that is not in a salt state, ethylchloroformate is reacted in an organic solvent (e.g., toluene) to obtain an intermediate step in the methyl removal reaction. Phosphorous ethyl 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine-1-carboxylate can be obtained. Ethyl 4-obtained (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine-1-carboxylic rate basic environment shifted by reacting in (for example, KOH aqueous solution) It is possible to obtain 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine from which methyl groups have been removed from roheptadine.

Step (b) is 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine R ₁ -Br, R obtained in step (a) depending on the desired substituent. _This is a step of reacting ₂ -Cl, R ₃ -COOH or R ₄ -COCl.

In Formula 1, when X is R ₁ , nucleophilic substitution of 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and alkyl bromide R ₁ -Br To react. That is, 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine, alkyl bromide (alkyl bromide) and K ₂ CO ₃ are organic solvents (eg, The target compound can be obtained by dissolving in THF) to react.

In Formula 1, when X is R ₂ , a nucleophilic substitution reaction of 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and the alkyl chloride R ₂ -Cl Order. That is, potassium carbonate, sodium iodide and alkyl chloride (alkyl chloride) are organic solvents of 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine (for example, DMF) The desired compound can be obtained by adding to the solution and reacting.

In Formula 1, when X is COR ₃ , amide coupling reaction of 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and the carboxylic acid R ₃ -COOH Order. That is, carboxylic acid, 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC), 4-(dimethylamino)pyridine (DMAP) and triethylamine are 4-(5 H -dibenzo[ The desired compound can be obtained by adding and reacting a,d ][7]anulen-5-ylidene)piperidine with an organic solvent (eg, dichloromethane) solution.

When X is COR ₄ in Chemical Formula 1, 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and acid chloride (carboxylic acid chloride) R _4- COCl is subjected to a nucleophilic substitution reaction. That is, an organic solvent (for example, dichloromethane) solution of 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine with carboxylic acid chloride and triethylamine The target compound can be obtained by adding and reacting with.

The present invention also provides a composition for promoting appetite in mammals comprising the compound as an active ingredient. As a result of the treatment of the ciproheptadine derivative of the present invention with the Kroe mHypoE cell line, it is expected to show an excellent appetite promoting effect by showing an AgRP (Agouti-related peptide) expression level similar to or lower than that of ciproheptadine. In addition, it was found that the compound exhibits an excellent effect of improving weight gain in broilers and piglets.

In one embodiment, the composition for promoting appetite may be a pharmaceutical composition, an animal medicine, or a feed additive. In one embodiment, the pharmaceutical composition may further include one or more pharmaceutically acceptable carriers or additives. That is, the pharmaceutical composition of the present invention can be formulated in the form of oral dosage forms, external preparations, suppositories, and sterile injection solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. . The pharmaceutically acceptable carriers include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants, are included. Oral solid preparations include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient, such as starch, calcium carbonate, sucrose or lactose ( lactose), gelatin, and the like, and may include a lubricant such as magnesium stearate and talc. Liquid formulations for oral use include suspensions, liquid solutions, emulsions, syrups, etc., and may include diluents such as water and liquid paraffin, wetting agents, sweeteners, fragrances, and preservatives. Parenteral preparations include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents, suspensions include vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, ethyl And injectable esters such as oleate. As a base for suppositories, witepsol, macrogol, tween, cacao butter, laurin butter, and glycerogelatin may be used.

The dosage of ciproheptadine derivative contained in the pharmaceutical composition of the present invention varies depending on the patient and animal condition and body weight, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by experts in the field. have. For example, the ciproheptadine derivative may be administered at a dose of 0.0001 to 1000 mg/kg per day, preferably 0.01 to 1000 mg/kg, and the administration may be administered once or several times a day. have. In addition, the pharmaceutical composition of the present invention may include the ciproheptadine derivative in an amount of 0.001 to 50% by weight based on the total weight of the composition.

The pharmaceutical composition of the present invention can be administered to a variety of routes to mammals such as rats, mice, livestock, and humans, for example, for oral, intraperitoneal, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater, or intracerebroventricular injection. It can be administered by.

The present invention also provides a method for promoting appetite in a mammal and a method for treating anorexia in a mammal.

Hereinafter, the present invention will be described in more detail through examples and test examples. However, the following examples and test examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.

<Example>

1. Structural design of ciproheptadine derivatives

In the present invention, a ciproheptadine derivative having histamine H1 receptor affinity was designed through computer molecular modeling and candidates were derived through synthesis (Examples 1 to 20). For the designed proheptadine derivative (20), Maestro 2017-1 (Schordinger) was used as a program for computer molecular modeling, and the protein 5-HT2B receptor (Code No. 5TVN) was used. Docking was attempted using this method.

Table 1 below shows the results of affinity analysis between the compound and histamine H1 receptor through molecular modeling.

Example Compound code Combined score rescue One CYP001 -7.286

2 CYP002 -8.196

3 CYP003 -7.486

4 CYP004 -7.992

5 CYP005 -6.949

6 CYP006 -7.773

7 CYP007 -4.902

8 CYP008 -6.169

9 CYP009 -5.725

10 CYP010 -6.386

11 CYP011 -5.686

12 CYP013 -6.627

13 CYP014 -

14 CYP015 -5.806

15 CYP016 -4.450

16 CYP018 -6.135

17 CYP019 -

18 CYP020 -6.413

19 CYP021 -5,557

20 CYP022 -5.875

As a result of molecular modeling, it was predicted that the compounds CYP002, CYP006, and CYP022 would exhibit the highest affinity for the histamine H1 receptor.

2. Synthesis of Ciproheptadine Derivatives

To introduce various substituents instead of the methyl group of nitrogen as a ciproheptadine derivative, demethylation and N -alkylation were attempted.

(1) Demethylation

[Level 1]

Method: Ciproheptadine hydrochloride sesquihydrate (5 g, 15.44 mmol, 1.0 eq.) was taken with 50 mL EtOAc and 10% aq. Basified with NaOH (20 mL). The organic layer was washed with water and dried over Na ₂ SO ₄ . The solvent was concentrated in vacuo to give free ciproheptadine. Ethylchloroformate (4.8 mL, 51.1 mmol, 3 eq.) was added to a solution of free ciproheptadine in toluene (100 mL). The reaction mixture was refluxed for 3 hours. The progress of the reaction was monitored by TLC using 10% methanol in dichloromethane as the mobile phase. The solvent was concentrated under vacuum to give an oily residue which was dissolved in EtOAc. The organic layer was washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to afford the desired compound as a white solid (4.8 g, 90%).

[Step 2]

By: Ethyl 4 in ethanol (80 mL) (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine-1-carboxylate [ethyl 4- (5 H - dibenzo[ a , d ][7]annulen-5-ylidene)piperidine-1-carboxylate] (4.8 g, 13.9 mmol, 1 eq.) and KOH (12.9 g dissolved in 20 mL of H ₂ O, 208 mmol, 15 Equiv.). The mixture was refluxed for 48 hours. The progress of the reaction was monitored by TLC using 10% methanol in dichloromethane as the mobile phase. The solvent was concentrated and the obtained residue was dissolved in ethyl acetate (EtOAc). Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to afford the desired compound as a white solid (3.5 g, 92%).

(2) Nucleophilic substitution reaction of alkyl bromide

Method: 4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine [4- (5 H -Dibenzo [a , d] [7] annulen-5-ylidene piperidine] (100 mg, 0.366 mmol, 1.2 eq), alkyl bromide (alkyl bromide) (0.305 mmol, 1.0 eq) and K ₂ CO ₃ (84 mg, 0.610 mmol, 2.0 eq) in THF (3 mL) Was added. The mixture was stirred at room temperature for 24 hours. The solvent was concentrated and the obtained residue was dissolved in ethyl acetate (EtOAc). Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo to give a crude compound, which was flash column chromatography using EtOAc/hexane (1/2) as eluent. purified by chromatography). The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product.

Table 2 below shows the structure and yield of a ciproheptadine derivative using an alkyl bromide.

(3) Nucleophilic substitution reaction of alkyl chloride

Method: Potassium carbonate (101 mg, 0.732 mmol, 2.0 eq), sodium iodide (82 mg, 0.549 mmol, 1.5 eq) and alkyl chloride (alkyl chloride) (0.366 mmol, 1.0 eq) were 4-(5 H -dibenzo [ a,d ][7]anulen-5-ylidene)piperidine (100 mg, 0.366 mmol, 1.0 eq) was added to a DMF (3.0 mL) solution. The mixture was stirred at 100 °C overnight. The solvent was concentrated and the obtained residue was dissolved in ethyl acetate (EtOAc). Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/2) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product.

Table 3 below shows the structure and yield of the ciproheptadine derivative using an alkyl chloride.

(4) Amide coupling reaction

Method: carboxylic acid (0.439 mmol, 1.2 eq), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide [1-(3-dimethylaminopropyl)-3-ethyl carbodiimide] (EDC, 105 mg, 0.549 mmol, 1.5 eq), 4-(dimethylamino)pyridine (DMAP, 4.5 mg, 0.037 mmol, 0.1 eq) and triethylamine (TEA, 74 mg, 0.732 mmol, 2.0 eq) 4-(5 H -dibenzo [ a,d ][7]anulen-5-ylidene)piperidine (100 mg, 0.366 mmol, 1.0 eq) was added to a dichloromethane (3.0 mL) solution. The mixture was stirred at 100 °C overnight. The solvent was concentrated and the obtained residue was dissolved in dichloromethane. Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/2) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product.

Table 4 below shows the structure and yield of the ciproheptadine derivative using carboxylic acid.

(5) Nucleophilic substitution reaction of acid chloride

Method: Carboxylic acid chloride (0.333 mmol, 1.0 eq) and triethylamine (TEA, 42 mg, 0.416 mmol, 1.25 eq) were prepared with 4-(5 H -dibenzo[ a,d ][7]anulen-5 -Ylidene)piperidine (100 mg, 0.366 mmol, 1.1 eq) was added to a dichloromethane (3.0 mL) solution. The mixture was stirred at room temperature overnight. The solvent was concentrated and the obtained residue was dissolved in dichloromethane. It was washed with 1N HCl. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/2) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product.

Table 5 below shows the structure and yield of the ciproheptadine derivative using acid chloride.

3. Structural analysis of cyproheptadine derivatives

The nuclear magnetic resonance spectroscopy (NMR) used for the analysis was performed using “ADVANCE III 400 NMR spectrometer” from “Bruker”, and was analyzed using a CDCl ₃ solvent containing TMS (tetramethylsilane) as a reference material. The analysis results are shown in FIGS. 1 to 20.

4. Evaluation of the activity of ciproheptadine derivatives (Test Example 1)

(1) Confirmation of change in AgRP expression level by ciprofeptapdine in mHypoE cell line

Compared with the control group treated with cyproheptadine on the Kroe mHypoE cell line (cellutions biosystems Inc.), it was confirmed that it has an effect of reducing the amount of AgRP (Agouti-related peptide) expression (see FIG. 21).

(2) Evaluation of AgRP expression change for ciproheptadine (CYP) derivatives in mHypoE cell line

As a result of the primary screening, 10 candidate groups (CYP001, CYP002, CYP003, CYP006, CYP008, CYP009, CYP010, CYP014, CYP015, CYP022) with a fold value of 0.5 or less when compared to the control among the CYP derivatives 5 candidate substances (CYP002, CYP006, CYP008, CYP014, CYP022) among these candidate groups were selected and repeated experiments were performed (see FIG. 22).

(3) Repeat experiment of AgRP expression level for 5 compounds of cyproheptadine (CYP) derivative selected from mHypoE cell line

As a result of repeated experiments using 5 selected candidate substances (CYP002, CYP006, CYP008, CYP014, CYP022), it was confirmed that the cells treated with CYP008 and CYP022 showed lower AgRP expression than the comparative group (see FIG. 23). .

CYP002, CYP006, and CYP022 were selected as lead materials for animal experiments considering the structural diversity and physical properties (solid) of the compounds. These three species also showed good affinity with the target protein in molecular modeling prediction results.

5. Scale-up synthesis of 3 kinds of lead substances (CYP002, CYP006, CYP022)

(1) CYP002

Method: 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine (10 g, 36.58 mmol, 1.2 eq.), 1-bromo-3-phenylpropane ( 6.17 g, 31.0 mmol, 1.0 eq) and K ₂ CO ₃ (8.57 g, 62.0 mmol, 2.0 eq) were added in THF (120 mL). The mixture was stirred at room temperature for 24 hours. The solvent was concentrated and the obtained residue was dissolved in ethyl acetate (EtOAc). Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/3) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product (7.0 g, 60%).

(2) CYP006

Method: 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine (8.20 g, 30.0 mmol, 1.2 eq.), 1-bromo-nonane [1-Bromo -nonane] (5.20 g, 25.0 mmol, 1.0 eq) and K ₂ CO ₃ (6.90 g, 50.0 mmol, 2.0 eq) were added in THF (120 mL). The mixture was stirred at room temperature for 24 hours. The solvent was concentrated and the obtained residue was dissolved in ethyl acetate (EtOAc). Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/2) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product (5.7 g, 48%).

(3) CYP022

Method: Phenylpropiolic acid (2.06 g, 14.08 mmol, 1.1 eq), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (EDC, 3.68 g, 19.21 mmol, 1.5 eq) , 4-(dimethylamino)pyridine (DMAP, 0.16 g, 1.28 mmol, 0.1 equiv) and triethylamine (TEA, 2.59 g, 25.61 mmol, 2.0 equiv) 4-(5 H -dibenzo[ a,d ] [7] Ananulene-5-ylidene)piperidine (3.50 g, 12.80 mmol, 1.0 eq.) was added to a dichloromethane (17.0 mL) solution. The mixture was stirred at room temperature overnight. The solvent was concentrated and the obtained residue was dissolved in dichloromethane. Then, it was washed with an appropriate amount of H ₂ O and brine. Finally, the organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give the crude compound, which was purified by flash column chromatography using EtOAc/hexane (1/2) as eluent. The desired fractions were collected and the solvent was evaporated on a rotary evaporator to give the product (3.19 g, 62%).

6. Qualitative analysis of three leading substances (CYP002, CYP006, CYP022)

The following apparatus and conditions were used to establish the analysis conditions of the three leading substances.

(Ⅰ) Apparatus: High Performance Liquid Chromatograph (HPLC), YL-9100 (Younglin Equipment)

(Ii) Analysis conditions

-Column: TC-C ₁₈ (Agilent, 4.6 mm × 250 mm, 5.0 μm)

-Mobile phase: A (0.1% Formic acid solution) / B (MeOH)

-Flow rate: 1.0 mL/min

-Column temperature: 40 ℃

-Detector: UV 254 nm

-Injection volume: 5 μL

The results of analyzing each material under the established conditions are shown in FIGS. 24 to 27.

<Experimental Example 2> Broiler weight increase experiment of a ciproheptadine derivative

1. Summary

This test was conducted to confirm the effect of new compounds on broiler growth.

1) Weight change

Cyproheptatine (positive control), CYP002, CYP006, and CYP022 per ton of feed was mixed with 1 kg each, and fed for 28 days to confirm the change in weight gain of each test group.

The total gain over the 28-day period of the negative control, positive control, CYP002, CYP006 and CYP022 administration groups prior to drug administration was 1129.40±112.64 g/bird, 1219.20±62.06 g/bird, 1147.40±91.88 g/bird, 1150.20±106.51 g/bird and 1256.80 ±101.35 g/bird, daily gains of 40.34±4.02 g/bird/day, 43.54±2.22 g/bird/day, 40.98±3.28 g/bird/day, 41.08±3.80 g/bird/day and 44.89±3.62 g It was /bird/day.

There was no significant difference between each test group in total weight gain and daily weight gain, but the weight gain of the CYP022 group was increased by about 11% compared to the negative control group, and the weight gain of the positive control group was increased by about 8% compared to the negative control group. The weight gain of the CYP002 and CYP006 groups was not different from that of the negative control group.

Even in the weekly gain, the weekly gain in the positive control and CYP022 groups was 14.09±2.35 g/bird/week and 14.34±0.87 g/bird/week, respectively, compared to the untreated group (11.11±2.87 g/bird/week). The weekly weight gain of the positive control group and the CYP022 group increased by about 12% compared to the negative control group.

2) Feed intake and feed conversion (FCR)

The 28 day negative control, positive control, CYP002, CYP006 and CYP022 administration groups confirmed the feed intake, 1876.07 g/bird, 1860.64 g/bird, 1886.69 g/bird, 1871.40 g/bird and 1866.86 g/bird, respectively. The feed conversion ratios (FCR) were 1.67±0.17, 1.53±0.08, 1.65±0.13, 1.64±0.15 and 1.49±0.12, respectively. Although there was no significant difference between the feed intake and feed conversion rates in each test group, the feed control rates of the positive control group and the CYP022 administration group tended to be lower than those of the negative control group, the CYP002 administration group and the CYP006 administration group.

3) Cortisol concentration change in blood

In order to confirm the effect of the new compound on blood cortisol, the cortisol concentration in the negative control group, positive control group, CYP002, CYP006 and CYP022 administration groups was 4.14±0.58 ng/mL, 3.76±0.64 ng, respectively. /mL 3.86±0.40 ng/mL, 3.88±0.54 ng/mL and 3.58±0.37 ng/mL, and no significance was observed between groups. However, the concentration of cortisol in the blood of all the test groups administered with the test substance tended to be lower than that of the negative control group.

Among the new compounds having a chemical structure similar to that of ciproheptadine, the CYP022 28-day gain was similar to that of the positive control, increased by 11%, respectively, compared to the negative control, and the blood coritosol concentration was also confirmed to be lower than that of the negative control. It was judged to be effective in improving the weight gain.

2. Purpose

This test was conducted to confirm the effect of adding new compound feed additives to broilers.

3. Materials

1) Cyproheptadine (positive control) [Fig. 28(a)]

2) CYP 002: Cyproheptadine derivative [Fig. 28(b)]

3) CYP 006: Cyproheptadine derivative [Fig. 28(c)]

4) CYP 022: Cyproheptadine derivative [Fig. 28(d)]

4. Test method

After obtaining approval through the deliberation procedure of the Hoseo University Animal Experimental Ethics Committee, animal experiments were performed (approval number: HSIACUC-18-167(1)).

1) Test system

-Lineage: broiler, Ross (1 day old)

-Supplier: Hana Bio Co., Ltd. (646, Hwaseo-dong, Paldal-gu, Suwon-si, Gyeonggi-do)

-Breeding environment: Temperature 22 ± 3 ℃, ventilation frequency 20 times / hour, lighting cycle 12 hours, illumination was set up using the animal laboratory of Hoseo University Safety Evaluation Center set to the environmental conditions of 30-40 Lux.

2) Composition of test group

The test group was configured as shown in Table 8 in order to confirm the weight increase effect in the broiler of the ciprohepdadine formulation feed additive. Fifteen to 20 broilers were stocked per test group, and 5 randomly selected on the day of stocking were used for the increase effect test.

3) Indicator of weight gain

-Body weight measurement: Body weight was measured twice a week to confirm the weight gain effect of the ciproheptadine preparation. The daily gain was determined by dividing the number of experimental days by the weight at the end of the test minus the weight at the start of vaccination.

-Measurement of feed intake: The feed was provided at 110 g per head, and the amount of feed intake was measured by measuring the daily supply and the remaining amount. The feed conversion ratio (FCR) was calculated by dividing the total feed intake by the total weight gain.

-Analysis of Cortisol concentration in blood: In order to analyze the effect of the test substance on the content of Cortisol in blood, Cortisol content in serum was analyzed using an ELISA kit (Bioassay Technology Laboratory). That is, 40 μL of serum, 10 μL of anti-cortisol antibody, and 50 μL of streptavidin-HRP were mixed in a well with chicken cortisol antibody attached and reacted at 37° C. for 60 minutes, and after the reaction was completed, washed 5 times with washing buffer Did. After adding 50 μL of Substrate A and B, respectively, the mixture was reacted at 37° C. for 10 minutes, and 50 μL of a stop solution was added to stop color development, and absorbance was measured at 450 nm using a UV spectrophotometer.

5. Results

1) Weight change

Before the drug administration, the test start weights of the negative control group, positive control group, CYP002, CYP006 and CYP022 administration groups were 43.80±2.49 g, 44.60±3.91 g, 45.40±4.62 g, 44.00±2.24 g and 45.00±3.94 g, respectively, and the end date of administration ( The weight of Day 28) was 1173.20±114.87 g, 1263.80±61.99 g, 1192.80±92.94 g, 1194.20±108.14 g and 1301.80±99.39 g, respectively. The total gain in each test group for 28 days was 1129.40±112.64 g/bird, 1219.20±62.06 g/bird, 1147.40±91.88 g/bird, 1150.20±106.51 g/bird, and 1256.80±101.35 g/bird, respectively, and the daily gain was 40.34. These were ±4.02 g/bird/day,43.54±2.22 g/bird/day, 40.98±3.28 g/bird/day, 41.08±3.80 g/bird/day and 44.89±3.62 g/bird/day. In total body weight and daily weight gain eopeotjiman significant difference of each test groups, were weight gain of CYP022 group is increased by about 11% as compared to the negative control, and body weight gain of the positive control group was increased by about 8% than the negative control group (Table 9, Fig. 30).

In addition, as a result of confirming the weekly weight gain of each test group, the 1 week weight gain of the positive control group and the CYP022 group was 14.09±2.35 g/bird/week and 14.34±0.87 g/bird/week, respectively, and the untreated group (11.11±2.87 g/ bird/week). However, in the weekly weight gain from Week 2 to Week 4, no significance was observed between the test groups, but in the week 4 weight gain, the weight gain in Week 4 of the positive control group and the CYP022 group increased by about 12% compared to the negative control group ( Table 9, Figure 31).

Table 9 below shows the weight, weekly weight gain, and daily weight gain for each test group.

2) Feed intake and feed conversion (FCR)

Each test substance was mixed with 1 kg per ton of feed, and fed freely for 28 days to check feed intake and feed efficiency.

The total feed intake of the negative control, positive control, CYP002, CYP006 and CYP022 administration groups was 1876.07 g/bird, 1860.64 g/bird, 1886.69 g/bird, 1871.40 g/bird and 1866.86 g/bird, respectively, and the feed intake of each group was similar. (Table 10).

The feed conversion rates (FCR) representing the productivity index of each test group were 1.67±0.17, 1.53±0.08, 1.65±0.13, 1.64±0.15, and 1.49±0.12, respectively, and there was no significance between the test groups. However, the feed conversion rate (FCR) of the positive control group and the CYP022 administration group was lower than that of the negative control group, the CYP002 administration group and the CYP006 administration group (Table 10). Table 10 shows feed intake and feed conversion of each test group.

3) Cortisol concentration in the blood

In order to analyze the effect of the new compound on the blood cortisol content, blood cortisol concentration was measured by taking blood at the end of the administration and as a result, blood cortisol concentrations in the negative control group, positive control group, CYP002, CYP006 and CYP022 administration groups were 4.14±0.58 ng/mL, respectively. , 3.76±0.64 ng/mL, 3.86±0.40 ng/mL, 3.88±0.54 ng/mL and 3.58±0.37 ng/mL, and no significance was observed between groups. However, the concentration of cortisol in the blood of all the test groups to which the test substance was administered tended to be lower than that of the negative control group (Table 11, FIG. 32).

Table 11 below shows the concentration of cortisol in the blood of each test group.

6. Discussion

Ciproheptadine is a first-generation H1 receptor antagonist and non-specifically acts as a 5-HT1A,5-HT2A receptor antagonist and is used as an appetite stimulant in people with reduced appetite due to various diseases such as gastrointestinal disorders and cystic fibrosis. When applied, it quickly absorbs and reaches the highest concentration in the blood within 3 to 4 hours, and the duration of action is known to be 6 to 12 hours. It easily passes through the BBB and acts on the central nervous system to act as an anticholinergic, antiserotonin, antihistamine, antidopamine, and sedative agent. It shows an effect.

Therefore, in the present invention, this test was performed to confirm the effect of broiler thickening of a new compound having a structure similar to that of ciproheptadine.

The total gain over the 28-day period of the negative control, positive control, CYP002, CYP006 and CYP022 administration groups prior to drug administration was 1129.40±112.64 g/bird, 1219.20±62.06 g/bird, 1147.40±91.88 g/bird, 1150.20±106.51 g/bird and 1256.80 There was no significant difference between each test group at ±101.35 g/bird, but the weight gain of the CYP022 group was increased by about 11% compared to the negative control group, and the weight gain of the positive control group was increased by about 8% compared to the negative control group. Although the feed intake of each test group was similar for 28 days, the feed conversion ratios (FCR) representing the productivity index were 1.67±0.17, 1.53±0.08, 1.65±0.13, 1.64±0.15, and 1.49±0.12, respectively. The conversion rate tended to be lower than that of the negative control, CYP002 and CYP006 groups.

Blood cortisol concentrations in the negative control, positive control, CYP002, CYP006 and CYP022 groups were 4.14±0.58 ng/mL, 3.76±0.64 ng/mL, 3.86±0.40 ng/mL, 3.88±0.54 ng/mL and 3.58±0.37 ng, respectively. /mL, and the concentration of cortisol in the blood of all the test groups administered with the test substance tended to be lower than that of the negative control group.

In conclusion, it was determined that CYP022 can be mixed with 1 kg per ton of feed to improve the amount of broiler chicken, etc., when fed daily to broilers.

<Test Example 3> Increased piglets of cyproheptadine derivatives

1. Summary

This test was carried out to confirm the effect of increasing the swine of new compounds. Of the 20 newly synthesized derivatives, three were selected in the laboratory and tested on piglets. As a product manufactured as a feed additive formulation, 1 kg per ton of feed was added to feed of piglets for 4 weeks.

Among the three selected new compounds (CYP002, CYP006, and CYP022), the total termination increase rate was 8.7% compared to the control group of CYP022, which showed a better growth effect than 4.8% of the positive control Cyproheptadine. This confirmed that it had a better weight increase effect than the existing substance, which is a positive control.

2. Purpose

This test was conducted to confirm whether feed additives using three new compounds have the effect of increasing piglets.

3. Materials

1) Cyproheptadine (positive control)

2) CYP 002: Cyproheptadine derivative

3) CYP 006: Cyproheptadine derivative

4) CYP 022: Cyproheptadine derivative

4. Test method

The experiment was conducted twice in consideration of the conditions of the test farm.

After obtaining the approval through the deliberation procedure of the Animal Experimental Ethics Committee of Kangwon National University, the animal experiment was conducted (approval number: KW-180822-1).

1) Test fund

-Piglets: weaned pigs (about 28 days old)

-Classification: 7 groups of 10 heads each (70 females), average weight of 5.0-6.0 kg.

-Breeding conditions and environment: The breeding density was early piglet (two/0.16 m2) and late piglet (two/0.31 m2), and after two weeks it was moved to late piglet and placed under the same conditions. The Donbang temperature setting was lowered by 1 degree every week at 28 degrees and stopped at 23 degrees, and the warming light was turned on until 1-4 weeks (the sensor was applied in the same way as the Donbang temperature setting, and turned ON and OFF).

2) Composition of test group

The test group was configured as shown in Table 12 and Table 13 in order to confirm the weight increase effect of the ciprohepdadine formulation feed additive. The test group composition of the first experiment (July 26-August 23, 2018) is shown in Table 12 below, and the test group configuration of the second experiment (August 30-2018 September 27, 2018) is shown in Table 13 below. Is shown.

3) Indicator of weight gain

-Body weight measurement: Body weight was measured to confirm the weight gain effect of ciproheptadine and three derivatives. Body weights were measured at the start and end of each test group.

-Measurement of feed intake: 1st week is fed 1+2 (mixed in a 1:1 ratio), 2nd week is fed 2nd. Week 3.4 was fed No. 3 feed, and the amount of feed consumed was measured by measuring the daily supply and balance. The feed conversion ratio (FCR) was calculated by dividing the total feed intake by the total weight gain.

-Termination increase rate by test group: It was calculated as the ratio of the total weight gain on the end day divided by the total weight on the start date.

-Completion of total increase rate of the control group compared to the control group: The value of the item of the total increase rate of the control group by the test group was determined as the difference between the total end increase rate of the control test groups in the first test and the second test.

5. Results

1) Weight change

As a result of the first test (July 26-August 23, 2018), the total weights of the test start dates of the negative control group, positive control group, and CYP022 administration group were 60.3±0.4kg, 60.3±0.5kg, and 48.5±0.5kg, respectively. , Body weight at the end of administration (Day 28) was 149.3±1.4kg, 152.2±1.6kg and 124.3±1.2kg. The control group, CYP group, and CYP022 group had 147.6%, 152.4%, and 156.3% of termination increase rates for each test group. As compared to the control group, the termination rate was 4.8% for the positive control (CYP) and 8.7% for the CYP022 (Table 14). The results of the first experiment (July 26-August 23, 2018) are shown in Table 14 below.

As a result of the 2nd trial (August 30, 2018-September 27, 2018), the total weight of the test initiation group of the control group, CYP002 and CYP006 administration group before administration of the drug was 54.0±0.5kg, 54.0±0.4kg, and 54.1±0.4kg, respectively. Body weights at the end day (Day 28) were 138.8±2.5 kg, 144.5±1.9 kg and 137.3±2.0 kg. The control group, CYP002 group, and CYP006 group had 157.0%, 167.6%, and 154% of the end-growth rate for each test group. Compared to the control group, the rate of termination increase was 10.6% in the CYP002 group and -3.2% in the CYP006 group (Table 15). Table 15 below shows the results of the second experiment (August 30, 2018-September 27, 2018).

2) Feed intake and feed conversion (FCR)

Each test substance was mixed with 1 kg per ton of feed, and fed freely for 28 days to check feed intake and feed efficiency.

In the first test, the total feed intakes of the negative control group, positive control group (CYP) and CYP022 group were 117.0kg, 122.2kg and 93.8kg, respectively (Table 16).

In the second test, the total feed intake of the negative control group, CYP002 and CYP006 administration group was 110.4kg, 111.9kg and 103.6kg, respectively.

Feed demand ratios representing the productivity index of each test group were 1.31, 1.33, and 1.33 in the first test, and 1.30, 1.24, and 1.25 in the second test, and the test group showed a lower tendency than the control (Table 16). Table 16 shows feed intake and feed conversion of each test group.

Table 17 below shows the results of comparing feed intake and weight gain.

As a result of comparing the feed intake of the administration group by the control group for each parking lot, the CYP group increased by 15% at week 1 and the CYP022 group increased by 13%. At Week 2, CYP increased 9% and CYP022 increased 0% (Table 17). Cyproheptadine and CYP022, which are the existing substances, showed a similar pattern in the first week, but four weeks of atrophy occurred in the CYP022 group (two in the other test group) as shown in the report of the farm at the second week. Eventually, due to umbilical cord hernia and hip dislocation, the experiment was terminated for the rest of the period, excluding two heads from CYP022. During the period of the 1st experiment, the overall experiment result was lower than the 2nd experiment due to the summer heat wave.

6. Discussion

Ciproheptadine is a first-generation H1 receptor antagonist and non-specifically acts as a 5-HT1A, 5-HT2A receptor antagonist, and is used as an appetite stimulant in people with reduced appetite due to various diseases such as gastrointestinal disorders and cystic fibrosis. When applied, it quickly absorbs and reaches the highest concentration in the blood within 3 to 4 hours, and the duration of action is known to be 6 to 12 hours. It easily passes through the BBB and acts on the central nervous system to act as an anticholinergic, antiserotonin, antihistamine, antidopamine, and sedative agent. It shows an effect.

Therefore, in the present invention, this test was performed to confirm the weight-increasing effect of piglets of a new derivative having a structure similar to that of ciproheptadine.

Due to the conditions of the experimental farm, the experiment was conducted in two. During the 1st experiment period (July 26 to August 23, 2018), it was confirmed that the increase in the effect of CYP022 was lower than the expected effect of the experiment in the laboratory due to the heat, but as a result, it was confirmed that the increase in the effect was higher than that of the existing compound, Cyproheptadine. . Also, feed intake increased by 15% and 13% in CYP and CYP022, respectively, at week 1 compared to the control.

In conclusion, it was confirmed that CYP022 was mixed with 1 kg per ton of feed to improve the amount of piglet gain when daily feeding to piglets.

Claims (13)

A compound of Formula 1, or a pharmaceutically acceptable salt thereof:
<Formula 1>

In the above formula, X is R ₁ , R ₂ , COR ₃ , or COR ₄ ,
R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ to C ₉ alkyl, substituted or unsubstituted C ₃ to C ₉ alkenyl, substituted or unsubstituted C ₃ to C ₉ alkynyl, or Dioxolanylalkyl,
R ₂ is alkoxybenzyl or alkylpyridine,
R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, phenylketonyl, or phenylalkynyl,
R ₄ is substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl. The compound according to claim 1, wherein R ₁ is benzyl, phenylpropyl, cinnamil, nonyl, allyl, butenyl, methylbutenyl, propynyl, or dioxolanylmethyl, or a pharmaceutically acceptable thereof. Possible salts. The compound according to claim 1, wherein R ₂ is methoxybenzyl or methylpyridine, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein R ₃ is cyclopentyl, cyclohexenyl, ethylacetyl, dihydroxyphenyl, methylphenylketonyl, or phenylethynyl, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein R ₄ is hexyl, tolyl, or benzyl, or a pharmaceutically acceptable salt thereof. The method of claim 1, wherein the compound
(1) 1-Benzyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine
(2) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-phenylpropyl)piperidine
(3) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(4-methoxybenzyl)piperidine]
(4) 1-cinnamyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine
(5) 3-((4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)methyl)pyridine
(6) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-nonylpiperidine
(7) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(prop-2-ain-1-yl)piperidine
(8) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)-1-(3-methylbut-2-en-1-yl)piperidine
(9) 1-allyl -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine
(10) (E) -1- (boot-2-en-1-yl) -4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidine
11 1 - ((1,3-dioxolan-2-yl) methyl) -4- (5 H-dibenzo [a, d] [7] annulene-5-ylidene) piperidine
(12) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (cyclopentyl) methanone
(13) 1- (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) octane-1-one
(14) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (cyclo hex-1-en-1-yl) -methanone
(15) Ethyl 3-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-oxopropanoate
(16) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (3,5-di-hydroxyphenyl) methanone
(17) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-2-phenylethane-1,2-dione
(18) (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl) (p- tolyl) methanone
(19) 1- (4- (5 H - dibenzo [a, d] [7] annulene-5-ylidene) piperidin-1-yl carbonyl) -2-phenyl-ethane-1-one
(20) 1-(4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidin-1-yl)-3-phenylprop-2-ain-1 -On
A compound, or a pharmaceutically acceptable salt thereof, characterized in that it is any one compound selected from the group consisting of. (a) synthesizing 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine by removing methyl group from ciprofheptane; And
(b) 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine and R ₁ -Br, R ₂ -Cl, R ₃ -COOH and R ₄ -COCl Reacting one selected from the group consisting of (where R ₁ is benzyl, phenylalkyl, cinnamil, substituted or unsubstituted C ₃ ~ C ₉ alkyl, substituted or unsubstituted C ₃ ~ C ₉ alkenyl , Substituted or unsubstituted C ₃ to C ₉ alkynyl, or dioxolanylalkyl; R ₂ is alkoxybenzyl or alkylpyridine; R ₃ is cycloalkyl, cycloalkenyl, alkylacetyl, dihydroxyphenyl, phenyl Ketonyl, or phenylalkynyl; R ₄ is substituted or unsubstituted C ₃ to C ₉ alkyl, tolyl, or benzyl). The method of claim 7, wherein in step (b), 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine is reacted with R ₁ -Br. Method of preparing a compound of Formula 1. The method of claim 7, wherein in step (b), 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine is reacted with R ₂ -Cl. Method of preparing a compound of Formula 1. The method of claim 7, wherein in step (b), 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine is reacted with R ₃ -COOH. Method of preparing a compound of Formula 1. The method of claim 7, wherein in step (b), 4-(5 H -dibenzo[ a,d ][7]anulen-5-ylidene)piperidine is reacted with R ₄ -COCl. Method of preparing a compound of Formula 1. A composition for promoting appetite in mammals comprising the compound of any one of claims 1 to 6 as an active ingredient. The composition for promoting appetite in mammals according to claim 12, wherein the composition for promoting appetite is an animal medicine or feed additive.

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