Composition for hardening soft tissue

Abstract

The present invention relates to various compounds capable of temporarily hardening soft tissue for surgical suturing of the soft tissue. The compounds according to the present invention can temporarily increase the hardness or tension of soft tissue, thereby improving the suturing efficiency during suturing of the soft tissue, thereby preventing aftereffects or the like from occurring due to insufficient anastomosis. In addition, the compounds according to the present invention can temporarily increase the hardness or tension of soft tissue, particularly pancreas, thereby increasing the suturing efficiency during pancreaticoduodenectomy and effectively preventing pancreatic leakage.

Claims

1. A compound or a pharmaceutically acceptable salt or hydrate thereof, wherein the compound is selected from the group consisting of the following compounds: 1) Cyclohexylmethyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; 2) 3-(tert-butoxy)-3-oxopropyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; 3) 2-fluorobenzyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; and 4) 2-(2-ethoxyethoxy)ethyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate.

2. A pharmaceutical composition for hardening soft tissue, comprising, as an active ingredient a compound according to claim 1.

3. The pharmaceutical composition of claim 2, wherein the soft tissue is one or more tissues selected from the group consisting of pancreas, liver, nerve, ligament, serosa, myofascia, intervertebral disc, and blood vessel.

4. A pharmaceutical composition for preventing pancreatic leakage, comprising, as an active ingredient a compound according to claim 1.

5. A method for preparing a compound according to claim 1, comprising the steps of: dissolving a compound represented by the following formula 10 and 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane) in an organic solvent; and adding an alkyl bromide to the dissolved solution, followed by stirring, thereby synthesizing a compound of claim 1: ##STR00071## wherein R is hydrogen or N(R′)(R″), and R′ and R″ are each independently hydrogen or a Ci-6 alkenyl group wherein an alkyl bromide is one or more selected from the group consisting of (bromomethyl)cyclohexane, tert-butyl bromopropionate, 2-fluorobenzyl bromide and 2-(2-ethoxyethoxy)ethyl bromide.

Description

DESCRIPTION OF DRAWINGS

(1) FIG. 1 graphically shows the changes in the hardness of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60027, KRM-60036, KRM-60037, KRM-60042 and KRM-60049 according to the present invention in one example of the present invention.

(2) FIG. 2 graphically shows the changes in the tension of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60027, KRM-60036, KRM-60037, KRM-60042 and KRM-60049 according to the present invention in one example of the present invention;

(3) FIG. 3 graphically shows the changes in the hardness of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60028, KRM-60029 and KRM-60030 according to the present invention in one example of the present invention.

(4) FIG. 4 graphically shows the changes in the tension of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60028, KRM-60029 and KRM-60030 according to the present invention in one example of the present invention.

(5) FIG. 5 graphically shows the changes in the hardness of the mouse pancreas after treatment of the pancreas with the antibiotics ICRP-00001 to KRP-000017 according to the present invention in one example of the present invention.

(6) FIG. 6 graphically shows the changes in the tension of the mouse pancreas after treatment of the pancreas with the antibiotics KRP-00001 to KRP-00017 according to the present invention in one example of the present invention.

(7) FIG. 7 graphically shows the changes in the hardness of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60077 to KRM-60088 according to the present invention in one example of the present invention.

(8) FIG. 8 graphically shows the changes in the tension of the mouse pancreas after treatment of the pancreas with the antibiotics KRM-60077 to KRM-60088 according to the present invention in one example of the present invention.

BEST MODE

(9) In accordance with one embodiment of the present invention, there is provided a compound represented by the following formula 1 or a pharmaceutically acceptable salt or hydrate thereof:

(10) ##STR00050##

(11) wherein R is hydrogen or N(R′)(R″); R′ and R″ are each independently hydrogen, a C.sub.1-6 alkyl group or a C.sub.1-6 alkenyl group; R.sub.1 is hydrogen or a C.sub.1-6 alkyl group; the C.sub.1-6 alkyl group of R.sub.1 is unsubstituted or substituted with R.sub.2; R.sub.2 is selected from the group consisting of a C.sub.1-6 alkenyl group, a C.sub.3-7 cycloalkyl group, a cyano group, a C.sub.1-6 alkyl group, a C.sub.6-10 aryl group, —OR.sub.3 and —COOR.sub.4; the aryl group of R.sub.2 is unsubstituted or substituted with halogen; R.sub.3 is —(CH.sub.2).sub.mOR.sub.5; m is an integer ranging from 0 to 3; and R.sub.4 and R.sub.5 are each independently a C.sub.1-6 alkyl group.

(12) In accordance with another embodiment of the present invention, there is provided a pharmaceutical composition for hardening soft tissue, comprising, as an active ingredient, the compound represented by formula 1 or a pharmaceutically acceptable salt or hydrate thereof.

(13) In accordance with still another embodiment of the present invention, there is provided a pharmaceutical composition for hardening soft tissue, comprising, as an active ingredient, a compound selected from the group consisting of a hydroxyquinoline compound, a derivative thereof, and a pharmaceutically acceptable salt and hydrate thereof.

(14) In accordance with still another embodiment of the present invention, there is provided a pharmaceutical composition for hardening soft tissue, comprising, as an active ingredient, a compound represented by any one of the following formulas 2 to 9 or a pharmaceutically acceptable salt or hydrate thereof:

(15) ##STR00051## ##STR00052##

MODE FOR INVENTION

(16) Hereinafter, the present invention will be described in detail with reference to examples. It will be obvious to those skilled in the art that these examples are only intended to illustrate the present invention in more detail and that the scope of the present invention as defined by the appended claims is not limited by these examples.

Example 1

Selection of Starting Compounds for Synthesis of Compounds for Hardening Pancreas

(17) To select starting compounds for synthesizing compounds that can enhance the function of the pancreas, a total of 23 antibiotics as shown below were prepared:

(18) ##STR00053## ##STR00054## ##STR00055##

(19) More specifically, three mice were treated with 1 mM of each of the antibiotics shown above, and three mice were treated with phosphate buffered saline (PBS) as a control. Furthermore, the tension and hardness of the pancreases of the mice treated with each antibiotic were compared with those of the pancreases of the control mice not treated with the antibiotic, and antibiotics showing increased tension and hardness upon treatment were selected as starting compounds.

(20) As a result, it was confirmed that among the 23 compounds, only KRM-60027, KRM-60036, KRM-60037, KRM-60042 and KRM-60049 compounds had the effects of increasing the tension and hardness of the pancreas. FIGS. 1 and 2 graphically show the changes in the hardness and tension of the pancreas after treatment with these five compounds compared to the control. Here, the hardness and the tension were measured using the methods described below.

(21) (1) Measurement of Hardness

(22) In order to measure the resistance of the pancreas using a hardness meter, the elastic modulus (EM) of the dissected pancreas was measured using Venustron (Axiom, Koriyama, Japan) based on the Hooke's law and the Hertz's contract stress theory, and the resistance of the pancreas was calculated using the following equation:

(23) $E=\frac{3}{4}.Math.F.Math.\left(1-{\upsilon }^2\right).Math.t^{-\frac{3}{2}}.Math.R^{-\frac{1}{2}}.$

(24) (F, force; v, Poisson's ratio; t, variant of compression; r, the diameter of a rigid spherical probe that presses an object vertically)

(25) (2) Measurement of Tensile

(26) The tension of the pancreas was measured using a dynamometer. Specifically, a pancreatic fragment having a size of 1×1×1.5 cm was prepared, and then silk 4-0 suture was passed through the tissue to form a knob, and suture-holding capacity was measured using a Newton dynamometer. At this time, the tension was calculated by adding the weight of the tissue.

(27) As shown in FIGS. 1 and 2, it could be seen that the pancreas of the mice was treated with each of the five antibiotics according to the present invention, that is, KRM-60027 (penicillin G potassium salt), KRM-60036 (ampicillin sodium salt), KRM-60037 (D-penicillamine), KRM-60042 (8-hydroxyquinoline) and KRM-60049 (ketoconazole), the tension and/or hardness of the pancreas significantly increased compared to the control. More specifically, it could be seen that KRM-60027 increased the tension of the pancreas to a significant level, and KRM-60036 significantly increased the hardness of the pancreas. In addition, it could be seen that KRM-60037, a drug having a simple structure and a molecular weight of only 149, exhibited the effect of increasing the tension of the pancreas by 1.5 times, and KRM-60042 and KRM-60049 increased both the hardness and tension of the pancreas to high levels.

(28) Based on these results, the five antibiotics were selected as starting compounds for compounds capable of hardening the pancreas.

Synthetic Example 1

Synthesis of Derivatives of KRM-60027

(29) The KRM-60027 compound which showed an excellent effect on an increase in the tension of the pancreas in Example 1 above was used as a starting material. Various alkyl bromide compounds were bound at the carboxylate position of KRM-60027, thereby synthesizing final compounds for enhancing the function of the pancreas.

Synthetic Example 1-1

Synthesis of KRM-60028

(30) ##STR00056##

(31) The KRM-60027 compound used as a starting material and 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane) were dissolved in DMF (dimethylformamide), and then 3-bromopropene as alkyl bromide was added thereto, followed by stirring at room temperature for 24 hours. After completion of the reaction was confirmed by thin-layer chromatography (TLC), ethyl acetate was added to the reaction solution which was then washed several times with water. The ethyl acetate layer containing the product was separated, and then anhydrous sodium sulfate was added thereto to remove the remaining water, followed by concentration. The concentrate was purified by column chromatography, thereby obtaining the final product (KRM-60028) in which the carboxylate of KRM-60027 was substituted with allyl.

Synthetic Example 1-2

Synthesis of KRM-60029

(32) ##STR00057##

(33) A final product in which the carboxylate of KRM-60027 was substituted with cyclohexyl was obtained in the same manner as described in Synthetic Example 1-1, except that 4-(bromomethyl)cyclohexane was used instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-3

Synthesis of KRM-60030

(34) ##STR00058##

(35) A final product in which the carboxylate of KRM-60027 was substituted with cyanobutyl was obtained in the same manner as described in Synthetic Example 1-1, except that 4-bromobutyronitrile was used instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-4

Synthesis of KRM-60077

(36) ##STR00059##

(37) A final product in which the carboxylate of KRM-60027 was substituted with tert-butyl propionate was obtained in the same manner as described in Synthetic Example 1-1, except that tert-butyl bromopropionate was used instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-5

Synthesis of KRM-60078

(38) ##STR00060##

(39) A final product in which the carboxylate of KRM-60027 was substituted with isobutyl was obtained in the same manner as described in Synthetic Example 1-1, except that 1-bromo-2-methylpropane was used instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-6

Synthesis of KRM-60079

(40) ##STR00061##

(41) A final product in which the carboxylate of KRM-60027 was substituted with 2-fluorobenzyl was obtained in the same manner as described in Synthetic Example 1-1, except that 2-fluorobenzyl bromide was used instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-7

Synthesis of KRM-60080

(42) ##STR00062##

(43) A final product in which the carboxylate of KRM-60027 was substituted with ethylene glycol was obtained in the same manner as described in Synthetic Example 1-1, except that 2-(2-ethoxyethoxy)ethyl bromide was used instead of 3-bromopropene as the alkyl bromide.

Example 2

Additional Selection of Compounds for Hardening Pancreas

(44) The pancreas of mice was treated with each of KRM-60028, KRM-60029 and KRM-60030 (synthesized in Synthetic Example 1 above) in the same manner as described in Example 1 above, and then changes in the hardness and tension of the pancreas compared to those of an untreated control group were measured. The results of the measurement are graphically shown in FIGS. 3 and 4.

(45) As shown in FIGS. 3 and 4, it could be confirmed that when the pancreas of mice was treated with each of KRM-60028, KRM-60029 and KRM-60030 according to the present invention, both the hardness and tension of the pancreas increased compared to those of the control. Referring to Example 1, the starting compound KRM-60027 showed an insignificant change in the hardness of the pancreas and a significant increase in the tension of the pancreas, compared to the control. It could be seen that KRM-60028, KRM-60029 and KRM-60030, which are compounds modified from KRM-60027, increased both the hardness and tension of the pancreas. In particular, the KRM-60030 compound is a derivative obtained by attaching a cyanopropyl group to the carboxylate group of KRM-60027, and has a nitrile group at the right end portion, and thus has a functional group capable of functioning as a hydrogen bonding acceptor. It could be seen that this KRM-60030 compound increase the hardness of the pancreas by 1.5 times and the tension of the pancreas by 2 times, compared to those of the control.

(46) This suggests that the compound represented by formula 1 according to the present invention can increase the hardness and tension of the pancreas, thereby temporarily hardening the pancreas.

Synthetic Example 2

Synthesis of Derivatives of KRM-60036

(47) The KRM-60036 compound which showed an excellent effect on an increase in the hardness of the pancreas in Example 1 above was used as a starting material. Alkyl bromide compounds were introduced selectively at the carboxylate position of KRM-60036 while maintaining the amine of KRM-60036, thereby synthesizing final compounds for enhancing the function of the pancreas. Specifically, as shown in reaction scheme 1 below, the amine of the KRM-60036 compound used as a starting material was subjected to boc protection. Then, as shown in reaction scheme 2 below, using 3-bromopropene, (bromomethyl)cyclohexane and 4-bromobutyronitrile as alkyl bromides, an alkyl group was introduced in the same manner as described in Synthetic Example 1-1 above, followed by boc deprotection, thereby obtaining final amine compounds.

(48) ##STR00063##

(49) ##STR00064## ##STR00065##

Example 3

Derivatives of KRM-60042

(50) The following derivative compounds were prepared, which are based on a hydroxyl group and quinoline which are the basic skeleton of the KRM-60042 compound (Sigma-Aldrich, cat. No 252565 148-24-3) which showed excellent effects on increases in both the hardness and tension of the pancreas in Example 1 above.

(51) ##STR00066## ##STR00067## ##STR00068##

(52) The pancreas of mice was treated with each of KRP-0001 to KRP-00017 (a total of 17 compounds) in the same manner as described in Example 1 above, and then the changes in the hardness and tension of the pancreas compared to those of an untreated control group were measured. The results of the measurement are graphically shown in FIGS. 5 and 6.

(53) As shown in FIGS. 5 and 6, it could be confirmed that among the 17 derivatives having the quinoline of KRM-60042 as a skeleton, treatment with 11 compounds (KRP-00001, KRP-00002, KRP-00003, KRP-00004, KRP-00005, KRP-00006, KRP-00009, KRP-00012, KRP-00013, KRP-00014 and KRP-00016) increased both the hardness and tension of the pancreas, and particularly, treatment with 10 compounds (ICRP-00001, KRP-00002, KRP-00003, KRP-00004, KRP-00005, KRP-00009, KRP-00012, KRP-00013, KRP-00014 and KRP-00016) increased the tension by at least 1.5 times, and treatment with KRP-00010 increased the tension by about 2.4 times, compared to that of the control group.

(54) This suggests that the hydroxyquinoline derivatives according to the present invention exhibited the effect of significantly increasing the tension of the pancreas and also increased the hardness of the pancreas.

Example 4

Other Compounds

(55) The pancreas of mice was treated with each of KRM-60077 to KRM-60080 (synthesized in Synthetic Example 1) and KRM-60081 to KRM-60088 compounds (shown below) in the same manner as described in Example 1 above, and then the changes in the hardness and tension of the pancreas compared to those of an untreated control group were measured. The results of the measurement are graphically shown in FIGS. 7 and 8.

(56) ##STR00069## ##STR00070##

(57) As shown in FIGS. 7 and 8, it could be confirmed that treatment with the KRM-60077 to KRM-60088 compounds increased the hardness and/or the tension of the pancreas compared to those of the control group, and particularly, treatment with the KRM-60077, KRM-60078, KRM-60080, KRM-60081, KRM-60082 and KRM-60088 compounds increased both the hardness and tension of the pancreas to significant levels.

(58) This suggests that the compounds according to the present invention exhibited the effect of significantly increasing the tension of the pancreas and also increased the hardness of the pancreas.

(59) The above experimental results suggest that when various modified compounds (derivatives) based on the structures of antibiotics that increase the tension and/or hardness of the pancreas are synthesized according to the present invention, compounds that effectively increase the function of the pancreas can be obtained. In addition, it can be seen that when the pancreas is treated with the compounds of the present invention, obtained as described above, the hardness and tension of the pancreas can temporarily increase, and thus the efficiency of suturing of the pancreas in surgical operations such as pancreaticoduodenectomy can increase, and furthermore, pancreatic leakage can also be prevented.

INDUSTRIAL APPLICABILITY

(60) The present invention relates to compounds capable to temporarily hardening soft tissue for surgical suturing of the soft tissue.