Biocompatible colloid preparation for promoting colorectal mucosa wound healing and application thereof

Abstract

The invention provides a biocompatible colloid preparation for promoting colorectal mucosa wound healing and application thereof.The colloid preparation comprises polyoxyethylene and water, the weight-average molecular weight of polyoxyethylene ranges from 1 million Dalton to 7 million Dalton, the weight-average molecular weight of polyoxyethylene ranges from 1 million Dalton to 1 million Dalton, and the weight-average molecular weight of polyoxyethylene ranges from 1 million Dalton to 1 million Dalton. The colloidal preparation is set to be capable of forming a protective colloidal mucosa when being applied to the surface of a colorectal wound mucosa, effectively protecting the colorectal mucosa and promoting wound healing.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine. Specifically, the present invention relates to a biocompatible colloidal preparation for promoting the healing of colorectal mucosal wounds. Background Art

[0002] Colonic ulcer is an acute or chronic inflammation occurring in the colonic mucosa and submucosa. After the occurrence of colonic ulcer, it causes local defects and ulcers. Common ulcer diseases mainly include: Ulcerative colitis (UC), Crohn's disease (CD), Ischemic colitis, Colonic tumor, Drug-induced enteritis, and Infective enteritis. Digestive tract diseases such as colon cancer and colonic obstruction formed ultimately according to its development often need to be treated by Colectomy.

[0003] Due to many reasons such as surgery, injury, inflammation, and infection, the intestinal mucosa of patients is damaged to form colonic ulcers, and even enterocutaneous fistulas. In the treatment of colonic ulcer diseases, the following several methods are mainly adopted clinically at present: (1) Intestinal fluid drainage. Using a double cannula for continuous negative pressure drainage to drain the leaked intestinal fluid to the outside of the body as much as possible. This method requires repeated drainage for 1 - 4 weeks, with a long healing time and increased pain for patients. (2) Using a medical adhesive to seal the enterocutaneous fistula. The adhesives currently used are mainly some compound preparations, such as compound zinc oxide ointment, etc. However, if the mucosal damage at the affected area is severe, in the case of ulcers and erosive surfaces, the effect of this adhesive is not significant, and it is also necessary to continuously deliver supplementary medicine through an anoscope after surgery to achieve the treatment effect, which also increases the pain of patients.

[0004] Based on the high incidence and severity of the above-mentioned intestinal mucosal damage, there is an urgent need in this field for a biocompatible colloid with good adhesiveness, capable of isolating the alkaline enzyme environment, and promoting the healing of colonic ulcers, for the treatment of colonic ulcers and the formation of enterocutaneous fistula closure. Summary of the Invention

[0005] In order to solve the problems brought by colonic ulcers and provide an effective repair and protection barrier for colorectal mucosal wounds, one of the purposes of the present invention is to provide a biocompatible colloidal preparation for promoting the healing of colorectal mucosal wounds, which contains polyethylene oxide and water, wherein:

[0006] The weight-average molecular weight of the polyethylene oxide is from 1 million Daltons to 7 million Daltons;

[0007] The colloidal preparation is configured to be capable of forming a protective colloidal mucosa when applied to the traumatized mucosal surface of the colorectum.

[0008] In some embodiments of the present invention, the raw materials of the colloidal preparation include polyoxyethylene particles with an average particle size of 0.5 μm to 2000 μm.

[0009] In some embodiments of the present invention, in the colloidal preparation, the ratio of polyoxyethylene to water ranges from 1 g / 50 mL to 1 g / 200 mL.

[0010] In some embodiments of the present invention, the fluidity of the colloidal preparation is suitable for delivery by colonoscopy.

[0011] In some embodiments of the present invention, the adhesion force of the protective colloidal mucosa to the intestinal mucosa is not less than 1 N.

[0012] In some embodiments of the present invention, the colloidal preparation takes polyoxyethylene particles as the main component of the preparation and further contains sodium carboxymethylcellulose.

[0013] In some embodiments of the present invention, the mass ratio of polyoxyethylene to sodium carboxymethylcellulose is 9:1 to 5:5.

[0014] In some embodiments of the present invention, the weight-average molecular weight of the polyoxyethylene is 2 million to 4 million daltons.

[0015] In some embodiments of the present invention, the colloidal preparation forms a protective colloidal mucosa when applied to the surface of the colonic mucosa.

[0016] On the other hand, another object of the present invention is to provide a medical use of the colloidal preparation according to the above various embodiments of the present invention, that is, to provide the application of the biocompatible colloidal preparation in the preparation of a drug for promoting the healing of colorectal mucosal wounds.

[0017] The present invention has the following advantages over the prior art: By selecting polyoxyethylene particles with a specific molecular weight and a specific particle size, and forming a colloidal preparation with water, the colloidal preparation can form a protective colloidal mucosa when applied to the traumatized mucosal surface of the colorectum, thereby effectively protecting the colorectal mucosa and promoting wound healing. In addition, the colloid formed by polyoxyethylene and water can be delivered to the rectum and colon by injection through a digestive endoscope, avoiding the problem of nozzle blockage caused by the nozzle sticking to mucus when polyoxyethylene is sprayed in the form of powder particles. Therefore, the colloidal preparation of the present invention is convenient for administration and the patient experience is better.

[0018] The present invention will be further described in detail below in conjunction with the drawings and specific embodiments. Brief Description of the Drawings

[0019] Figure 1 It is a schematic diagram of the adhesion force test used to evaluate the adhesion performance in the embodiments of the present invention;

[0020] Figure 2 It is a schematic diagram of the flushing experiment simulating the colon environment in the embodiments of the present invention;

[0021] Figure 3 They are colonoscopy photos of the porcine colon mucosa wound before and four weeks after applying the colloidal preparation #2 in Example 1 and the control group. Detailed implementation manners

[0022] The technical solutions of the present invention will be further described below in conjunction with the exemplary embodiments and the accompanying drawings of the present invention. Obviously, the embodiments described herein are only for illustrative purposes and do not exhaust all embodiments of the present invention. The following description of the exemplary embodiments is merely illustrative and does not constitute any limitation to the protection scope of the present invention. Based on the embodiments of the present invention, all other implementation manners obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0023] Unless otherwise specifically stated, the numerical values and numerical ranges recorded in the exemplary embodiments of the present invention do not constitute any limitation to the protection scope of the present invention. For the sake of brevity, the technologies and methods known to those of ordinary skill in the relevant fields are not described in detail herein, but in appropriate cases, the known technologies and methods should be regarded as part of this specification. In all examples recorded herein, any specific value should be construed as merely exemplary and not constituting any limitation. Therefore, other examples except the exemplary embodiments may have different values.

[0024] Example 1

[0025] This embodiment provides colloidal preparations #1-#4 for colon mucosa protection, which respectively contain polyoxyethylene (PEO) with different weight average molecular weights.

[0026] Specifically, the colloidal preparations of this embodiment are prepared according to the following method:

[0027] (a) Add the raw material PEO particles of each preparation according to Table 1;

[0028] (b) Add purified water according to the ratio of 1 g:100 ml, stir until a colloid is formed, thereby obtaining colloidal preparations #1-#4.

[0029] Table 1 Physical and chemical parameters of the PEO raw materials for preparing colloidal preparations #1-#4

[0030] Formulation number Physicochemical parameters of PEO #1 Weight-average molecular weight: 1 million; Particle size: 0.5 μm to 2000 μm; #2 Weight-average molecular weight: 2 million; Particle size: 0.5 μm to 2000 μm; #3 Weight-average molecular weight: 4 million; Particle size: 0.5 μm to 2000 μm; #4 Weight-average molecular weight: 7 million; Particle size: 0.5 μm to 2000 μm;

[0031] When the colloidal preparations #1 - #4 are applied to the traumatized mucosal surface of the colorectum, they can form a protective colloidal mucosa, thus effectively protecting the colorectal mucosa and promoting wound healing.

[0032] To evaluate the protective and repair-promoting effects of the colloidal preparations #1 - #4 of this example on the colonic mucosa, the adhesion force of each colloidal preparation to the colonic mucosa, the adhesion effect on the colonic mucosa, as well as the effects on promoting colonic mucosal healing and mucosal protection were tested respectively as follows.

[0033] 1.1 Adhesion force test on the colonic mucosa

[0034] 1.1.1 Test purpose:

[0035] Evaluate the adhesion performance of the colloidal preparations #1 - #4 to the colonic mucosa through the adhesion force test.

[0036] 1.1.2 Test method:

[0037] Use the Figure 1 shown tester for the test. Take the excised porcine colon and lay its mucosal layer out flat on the robotic arms at both ends of the adhesion force tester. Take 1 ml of each of the above-prepared colloidal preparations #1 to #4 and spread them evenly on the lower mucosal surface. Set the parameters of the tester as follows: compression speed: 20 mm / min, switching condition: force ≤ -50 gf, pause time: 0.5 min, stretching speed: 20 mm / min; start the tester and control the movement of the robotic arm according to the set parameters to test the adhesion force.

[0038] 2.1.3 Test results:

[0039] The adhesion forces of the colloidal preparations #1 to #4 to the colonic mucosa tested according to the above test method are shown in Table 2. As can be seen from Table 2, the adhesion forces of the protective colloidal mucosa formed by the colloidal preparations #1 to #4 to the intestinal mucosa are not less than 1 N.

[0040] Table 2 Test results of the adhesion force of the #1 - #4 preparations to the colonic mucosa

[0041] #1 #2 #3 #4 Adhesion force (N) 1.78 1.66 1.57 1.55

[0042] 1.2 Test on the adhesion effect on the colonic mucosa

[0043] 1.2.1 Test purpose:

[0044] In order to provide an effective repair and protection barrier for the colonic mucosal injury wound, the protective colloidal mucosa formed by the biocompatible colloidal preparation of the present invention on the colonic mucosa should last for a sufficient time.

[0045] The following is a simulation test on the duration of the colloidal preparations #1 to #4 in the colonic environment.

[0046] 1.2.2 Test method:

[0047] Adopt the Figure 2 inclined plate device as shown to simulate the colon environment for tolerance testing. Take the excised porcine colon, lay the mucosal layer outwards flat on the inclined plate device, take 1 ml of each of the above #1-#4 colloidal preparations and spread them on the surface of the colon mucosa, and drop in a staining agent for easy observation. Subsequently, rinse it with artificial intestinal fluid at a rate of 1500 ml / 24 h, and observe the residence time of the protective colloidal mucosa formed on the surface of the colon mucosa under the rinsing condition.

[0048] Among them, the preparation method of artificial intestinal fluid: Take 6.8 g of potassium dihydrogen phosphate, add 500 mL of water to dissolve it, adjust the pH value to 6.8 with 0.1 mol / L sodium hydroxide solution. Another 10 g of trypsin is weighed and dissolved in an appropriate amount of water. After mixing the two solutions, add water to make up to 1000 mL to obtain artificial intestinal fluid. 1.2.3 Test results:

[0049] Under this condition, each of the above #1-#4 colloidal preparations can adhere and stay on the surface of the colon mucosa for more than 72 hours.

[0050] Thus, it can be seen that the colloidal preparation has sufficient tolerance to artificial intestinal fluid and will not degrade and lose its adhesion performance due to the weak alkalinity in the colon. At the same time, it has sufficient adhesion performance to the colon mucosa, the adhesion force is greater than 1 N and it can stay on the mucosa surface for at least 72 hours. Usually, the colon mucosa can be repaired within a period not exceeding 72 hours. Therefore, the adhesion ability of the preparation of the present invention to the colon mucosa for not less than 72 hours in the colon environment can effectively isolate the damaged wound surface from the alkali and / or enzyme in the colon, thereby promoting the repair of colon mucosal damage.

[0051] 1.3 Test on promoting healing and mucosal protection effect of colon mucosa

[0052] 1.3.1 Test purpose:

[0053] Test the promoting healing effect of the above #1-#4 colloidal preparations on the colon through animal experiments.

[0054] 1.3.2 Method:

[0055] The experiment is carried out using Bama pigs. Select Bama pigs aged 3-6 months and with a body weight range of 40-45 kg, divided into 4 experimental groups and 1 blank control group, with 2 pigs in each group.

[0056] In order to determine the general health status of the animals, a baseline assessment is carried out on all experimental pigs, including blood routine, complete blood count, serum chemistry, blood glucose and liver and kidney functions. Then place the experimental pigs in the lateral position and perform endoscopic examination to observe changes such as bleeding, tumors, ulcers, inflammatory lesions, etc.

[0057] Anesthesia was performed by injecting sodium pentobarbital into the marginal ear vein or other appropriate methods. Subsequently, the experimental pigs were fixed supine on the operating table, tracheal intubation was performed, and assisted ventilation was provided by a ventilator. The vital signs of the animals (heart rate, blood pressure, pulse, respiratory rate, blood oxygen saturation, ET-CO2, etc.) were monitored throughout the process, and no abnormal vital signs were observed during the experiment. During the experiment, normal saline was intravenously injected into all experimental pigs to maintain the mean blood pressure above 60 mmHg.

[0058] Under colonoscopy, a snare was used for cold resection to form a wound surface of about 2 cm × 2 cm in the colon area, and the model was successfully established.

[0059] One hour after the wound surface was formed, 10 ml of the colloidal preparations #1-#4 were respectively injected once under colonoscopy in each experimental group, and the colloid was observed until the wound surface was completely covered. The control group was not injected with the colloidal preparation.

[0060] After the operation, the animals were normally raised for four weeks. Four weeks later, endoscopic follow-up was performed to observe the healing of the wound surface area and calculate the wound surface area.

[0061] 1.3.3 Test results:

[0062] After calculation, the colon mucosa wound surface areas of each experimental group and the control group four weeks later are shown in Table 3 below. It can be seen from Table 3 that the colon mucosa wound surface areas of each experimental group were significantly reduced and the healing effect was better, while the colon mucosa wound surface area of the control group was still large and the healing effect was not as good as that of the experimental group.

[0063] Figure 3 These are the colonoscopy photos of the porcine colon mucosa wound surface before and four weeks after applying the colloidal preparation #2 in Example 1 and the control group. It can be seen from this figure that most of the colon mucosa wound surface in the experimental group injected with the #2 colloidal preparation had healed four weeks after administration, and the damaged wound surface was hardly visible, while most of the colon mucosa wound surface of the experimental pigs in the control group had not healed four weeks later. Thus, it can be seen that the colloidal preparation of the experimental group can effectively promote the healing of the colon mucosa wound surface.

[0064] Table 3 Colon mucosa wound surface areas of each experimental group four weeks later

[0065] #1 #2 #3 #4 Control group <![CDATA[Ulcer area (cm 2 )]]> 1.6 1.4 1.7 1.5 3.2

[0066] Example 2

[0067] This example provides a series of mucosal protection colloids containing polyoxyethylene and sodium carboxymethylcellulose, namely colloidal preparations #6-#10, wherein:

[0068] Polyoxyethylene (PEO): weight-average molecular weight of 2 million, and the average particle size of its raw material particles is 0.5 μm to 2000 μm;

[0069] Sodium carboxymethyl cellulose: particle size 0.5 μm to 2000 μm, degree of substitution ≥ 9, viscosity 5000.

[0070] The colloidal preparations #6 - #10 are prepared according to the following steps:

[0071] (a) Add various raw materials according to the mass ratio of components in Table 4 and mix evenly;

[0072] (b) Add purified water according to the ratio of 1 g (polyethylene oxide + sodium carboxymethyl cellulose): 100 ml, stir until a colloid is formed to obtain the colloidal preparations #6 - #10.

[0073] Table 4 Composition ratio of the colloidal preparations #6 - #10

[0074] Formulation number Component mass ratio #6 Polyethylene oxide: Sodium carboxymethyl cellulose = 9:1 #7 Polyethylene oxide: Sodium carboxymethyl cellulose = 8:2 #8 Polyethylene oxide: Sodium carboxymethyl cellulose = 7:3 #9 Polyethylene oxide: Sodium carboxymethyl cellulose = 6:4 #10 Polyethylene oxide: Sodium carboxymethyl cellulose = 5:5

[0075] In order to evaluate the protective and repair - promoting effects of the colloidal preparations #6 - #10 of this example on the colonic mucosa, the same method as in Example 1 was used to test the performance of the prepared colonic mucosa - protecting colloidal preparations #6 - #10.

[0076] 2.1 Adhesion force test on the colonic mucosa

[0077] 2.1.1 Test purpose:

[0078] Evaluate the adhesion performance of the colloidal preparations #6 - #10 to the colonic mucosa through the adhesion force test.

[0079] 2.1.2 Test method:

[0080] Use Figure 1 the shown tester. Take the excised porcine colon, lay the mucosal layer outwards flat on the adhesion force tester, and evenly spread 1 ml of each of the above - prepared colloidal preparations #6 to #10 on the lower mucosal surface. Set the tester parameters: compression speed: 20 mm / min, switching condition: force ≤ - 50 gf, pause time: 0.5 min, stretching speed: 20 mm / min; start the tester and control the movement of the robotic arm according to the set parameters to test the adhesion force.

[0081] 2.1.3 Test results

[0082] The adhesion forces of the colloidal preparations #6 to #10 to the colonic mucosa tested according to the above test method are shown in Table 5. As can be seen from Table 5, the adhesion forces of the protective colloidal mucosa formed by the colloidal preparations #6 to #10 to the intestinal mucosa are all not less than 1.5 N.

[0083] Table 5 Test results of the adhesion force of the #6 - #10 preparations to the colonic mucosa

[0084] #6 #7 #8 #9 #10 Adhesion force (N) 1.67 1.69 1.89 1.75 1.69

[0085] 2.2 Test on the adhesion effect on colonic mucosa

[0086] 2.2.1 Test purpose:

[0087] To test the duration of the protective colloidal mucosa formed by the colloidal preparations #6 to #10 of this embodiment on the colonic mucosa and its tolerance to the colonic environment by simulating the colonic environment.

[0088] 2.2.2 Test method:

[0089] Prepare artificial intestinal fluid. Take 6.8 g of potassium dihydrogen phosphate, add 500 mL of water to dissolve it, adjust the pH value to 6.8 with 0.1 mol / L sodium hydroxide solution. Another 10 g of trypsin is weighed and dissolved in an appropriate amount of water. After mixing the two solutions, add water to make up to 1000 mL to obtain artificial intestinal fluid.

[0090] Same as Example 1, use the inclined plate device as shown in Figure 2 to test the tolerance by simulating the colonic environment. Take the excised porcine colon, lay the mucosal layer outwards on the inclined plate device, take 1 mL of each of the above-mentioned colloidal preparations #6 - #10 and spread it on the surface of the colonic mucosa, and drop in a staining agent for easy observation. Subsequently, rinse it with artificial intestinal fluid at a rate of 1500 mL / 24 h, and observe the residence time of the protective colloidal mucosa formed on the surface of the colonic mucosa under the rinsing condition.

[0091] 2.2.3 Test results:

[0092] Under this condition, the above-mentioned colloidal preparations #6 - #10 can all adhere and stay on the mucosal surface for more than 72 hours.

[0093] Thus, it can be seen that the colloidal preparations #6 - #10 have sufficient tolerance to artificial intestinal fluid, will not degrade and lose their adhesion performance due to the weak alkalinity in the colon, and at the same time they have sufficient adhesion performance to the colonic mucosa, with an adhesion force greater than 1 N and can stay on the mucosal surface for at least 72 hours.

[0094] 2.3 Test on the promoting healing and mucosal protection effects on colonic mucosa

[0095] 2.3.1 Test purpose:

[0096] To test the promoting healing effect of the above-mentioned colloidal preparations #6 - #10 on the colon through animal experiments.

[0097] 2.3.2 Test method:

[0098] Same as Example 1, the experiment was conducted in healthy Bama pigs. Bama pigs aged 3 - 6 months with a body weight range of 40 - 45 kg were selected and divided into 5 experimental groups and 1 blank control group, with 2 pigs in each group.

[0099] To determine the general health status of the animals, a baseline assessment was performed on all experimental pigs, including blood routine, complete blood count, serum chemistry, blood glucose, and liver and kidney functions. Then the experimental pigs were placed in the lateral decubitus position for endoscopic examination to observe any changes such as bleeding, tumors, ulcers, and inflammatory lesions.

[0100] Anesthesia was induced by intravenous injection of sodium pentobarbital into the marginal ear vein or other appropriate methods. Subsequently, the experimental pigs were fixed supine on the operating table, tracheal intubation was performed, and ventilation was assisted by a ventilator. The vital signs of the animals (heart rate, blood pressure, pulse, respiratory rate, oxygen saturation, ET - CO2, etc.) were monitored throughout the process, and no abnormal vital signs were observed during the experiment. During the experiment, normal saline was intravenously injected into all experimental pigs to maintain the mean blood pressure above 60 mmHg.

[0101] Under colonoscopy, a cold snare resection was used to form a wound surface of approximately 2 cm × 2 cm in the colon, and the model was successfully established.

[0102] One hour after the formation of the wound surface, 10 ml of the colloid preparations #6 - #10 were respectively injected once under colonoscopy in each experimental group, and the colloid was observed until it completely covered the wound surface. The control group was not injected with the colloid preparation.

[0103] After the operation, the pigs were normally raised for four weeks. Four weeks later, endoscopic follow - up was performed to observe the healing of the wound surface at the wound site, and the wound surface area was calculated.

[0104] 2.3.3 Test results:

[0105] There was residual colloid on the wound surfaces of the experimental groups. After the above calculation, the colon mucosa wound surface areas of each experimental group and the control group are shown in Table 6 below. The average ulcer area of the control group was 3.2 cm 2 . The average ulcer areas of the experimental groups #6 - #10 were respectively: 0.7 cm 2 , 1.1 cm 2 , 1.2 cm 2 , 0.9 cm 2 , 1.5 cm 2 . It can be seen from the data that after four - week healing after using the colloids #6 - #10 of the present invention, the wound surface area was significantly reduced compared with the control group, and the preparations of the present invention can all promote the healing of the intestinal mucosa injury wound surface.

[0106] Table 6 Colon mucosa wound surface conditions of each experimental group after four weeks

[0107] #6 #7 #8 #9 #10 Control group <![CDATA[Ulcer area (cm 2 )]]> 0.7 1.1 1.2 0.9 1.5 3.2

[0108] The present invention has been specifically described above in conjunction with specific embodiments. These specific embodiments are merely exemplary and cannot be used to limit the protection scope of the present invention. Those skilled in the art can make various modifications, changes or substitutions to the present invention without departing from the essence and scope of the present invention. Therefore, various equivalent changes made in accordance with the present invention still fall within the scope covered by the present invention.

Claims

1. A biocompatible colloidal preparation for promoting the healing of colorectal mucosal wounds, which comprises polyoxyethylene and water, wherein: The weight-average molecular weight of the polyoxyethylene is from 1 million Daltons to 7 million Daltons; The colloidal preparation is configured to be capable of forming a protective colloidal mucosa when applied to the traumatized mucosal surface of the colorectum.

2. The colloidal preparation according to claim 1, wherein The raw materials of the colloidal preparation include polyoxyethylene particles with an average particle size of 0.5 μm to 2000 μm.

3. The colloidal preparation according to claim 1, wherein in the colloidal preparation, the ratio of polyoxyethylene to water ranges from 1 g / 50 mL to 1 g / 200 mL.

4. The colloidal preparation according to claim 1, wherein the fluidity of the colloidal preparation is suitable for delivery by a colonoscope.

5. The colloidal preparation according to claim 1, wherein the adhesion force of the protective colloidal mucosa to the intestinal mucosa is not less than 1 N.

6. The colloidal preparation according to claim 1, further comprising sodium carboxymethylcellulose.

7. The colloidal preparation according to claim 6, wherein The mass ratio of polyoxyethylene to sodium carboxymethylcellulose is from 9:1 to 5:

5.

8. The colloidal preparation according to claim 1, wherein, The weight-average molecular weight of the polyoxyethylene is from 2 million Daltons to 4 million Daltons.

9. The colloidal preparation according to claim 1, wherein, The colloidal preparation forms a protective colloidal mucosa when applied to the surface of the colonic mucosa.

10. Use of the colloidal preparation according to any one of claims 1 to 9 in the preparation of a drug for promoting the healing of colorectal mucosal wounds.

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