A food-derived placebo for clinical trial of stroke-refreshing solution and its preparation method

Abstract

The invention discloses a food-based placebo for a clinical trial of a stroke brain-awakening liquid. The placebo is mainly prepared from the following raw materials: soy protein powder, gluten powder, protease, maltodextrin, brown sugar syrup, steviol glycoside, edible salt, monosodium glutamate, edible flavor, potassium sorbate, DL-malic acid and water. The invention also discloses a preparation method of the placebo. The placebo is completely prepared from food raw materials and is similar to a test drug in appearance, smell, taste and other aspects and is not easily recognized by researchers and subjects. The placebo can prevent bias and blind breaking in double-blind and controlled clinical trials to the greatest extent, thereby ensuring the quality of the clinical trials.

Description

Technical Field

[0001] The invention belongs to the technical field of placebos, and in particular relates to a food-derived placebo for clinical trials of stroke-revitalizing liquid and a preparation method thereof. Background Art

[0002] Traditional Chinese medicine has encountered practical difficulties in its promotion, mainly due to the lack of theoretical basis, clinical empirical research and mechanism of action, and its effectiveness is urgently needed to be scientifically proven. Control, randomization, blinding and repetition are the basic principles for the implementation of clinical trials of new drugs. Control is one of the methods to test whether there is a systematic error in the measurement method, and it is also an effective method to evaluate the effectiveness and safety of experimental drugs. Common control group settings in clinical trials include five types: positive drug control, placebo control, blank control, dose control and external control. Among them, placebo control has the function of removing the placebo effect and objectively distinguishing the efficacy and safety of the experimental drug, so it is widely used in confirmatory clinical trials.

[0003] Placebo became a medical term as early as the end of the 18th century, used to refer to something that can comfort and please patients despite having no effect. With the application of evidence-based medicine in the field of traditional Chinese medicine, the use of standardized clinical trials to evaluate the efficacy of traditional Chinese medicine has become increasingly important. In modern medical clinical research, the use of placebos has a long history and plays an important role in blind research. In order to implement the principle of blinding, the preparation technology of placebos is one of the keys to the implementation of blind trials. Placebos used as controls cannot contain test drugs. In addition to meeting the requirements of hygienic inspections, they are usually required to be consistent with the therapeutic drugs in appearance, smell, and color. The placebo effect refers to the impact of the patient's strong belief that he or she will be cured by this drug or method on the body. The use of placebo control is to exclude the placebo effect and to reflect the efficacy and safety of the drug to the greatest extent possible. In principle, under the premise of compliance with ethics, the clinical trial data of placebo control should be used to conduct risk / benefit assessment, and the importance of clinical value in risk / benefit assessment should be fully considered to evaluate whether it has market value.

[0004] There is no shortage of placebo applications in modern medical research. Placebos play an indispensable role in most large-scale randomized double-blind controlled studies. Western medicine placebos have a single ingredient and are relatively easy to make. However, Chinese medicine placebos are complex in composition. For some Chinese medicine preparations, the natural ingredients they contain can often be as high as hundreds or even more. Therefore, there are great difficulties in the production process of Chinese medicine placebos. Simulating the taste, smell, and appearance of Chinese medicine preparations is still a major problem in the field of Chinese medicine placebo production.

[0005] The stroke refreshing solution is composed of red ginseng, Panax notoginseng, raw rhubarb, etc., and the auxiliary materials are sorbic acid and polysorbate 80. The main treatment is to invigorate qi and refresh the brain. It is used for patients with acute cerebral hemorrhage and acute cerebral infarction. Red ginseng is used as the main drug to replenish vital energy, targeting the pathological essence of vital energy deficiency, and replenishing qi to promote blood circulation, strengthening the function of Panax notoginseng and other herbs in the prescription to promote blood circulation and remove blood stasis, so as to remove blood that has left the meridians. There is currently no placebo specifically for the stroke refreshing solution. Summary of the invention

[0006] The purpose of the present invention is to provide a food-derived placebo for clinical trials of stroke brain-awakening liquid, which is similar to the experimental drug stroke brain-awakening liquid in appearance, smell, taste, etc., and is not easily identified by researchers and subjects. It can prevent bias and blinding in double-blind, placebo-controlled clinical trials to the greatest extent, thereby ensuring the quality of clinical trials.

[0007] The present invention also aims to provide a method for preparing the food-derived placebo for clinical trials of the above-mentioned stroke-refreshing solution, which has a simple process.

[0008] The first object of the present invention is achieved by the following technical solution: a food-based placebo for clinical trials of stroke-refreshing solution, wherein the placebo is mainly made of the following raw materials in percentage by mass:

[0009]

[0010] The invention adopts the sensory characteristics of the stroke refreshing liquid, such as color, smell, taste, etc., avoids using raw materials with both medicinal and edible functions, and finally selects food-grade raw materials that are safe and reliable without medical value.

[0011] The food-derived placebo for the clinical trial of the stroke awakening solution of the present invention is prepared entirely from food raw materials and is a placebo that is similar to the test drug in appearance, smell, taste, etc. and is not easily identified by researchers and subjects. It can prevent bias and unblinding in double-blind, controlled clinical trials to the greatest extent possible, thereby ensuring the quality of the clinical trials.

[0012] Preferably, the mass ratio of the soy protein powder to the gluten powder is 3 to 6:1.

[0013] Preferably, the protease is one or more of papain, pepsin and trypsin.

[0014] Since the stroke refreshing liquid has a medicinal smell similar to that of Salvia miltiorrhiza and Chuanxiong, when choosing edible flavors, you can choose commercial food flavors with similar ginseng plant flavors, such as ginseng flavor.

[0015] The food-derived placebo for clinical trial of stroke awakening solution in the present invention is mainly prepared by the following steps: raw material pretreatment, enzymatic hydrolysis, separation, step-by-step preparation, mild reaction, sterilization, cooling and filling.

[0016] The second object of the present invention is achieved by the following technical solution: The preparation method of the food-derived placebo for the clinical trial of the stroke-refreshing solution comprises the following steps:

[0017] (1) Pretreatment: Mix the soybean protein powder and gluten powder evenly to obtain mixed soybean protein powder, and store the mixed soybean protein powder at 55 to 75° C. for 30 to 60 minutes;

[0018] (2) Enzymatic hydrolysis: Mix the mixed soybean protein powder, protease and water, stir, and perform enzymatic hydrolysis at 50°C to 66°C for 3 to 8 hours, then inactivate the enzyme;

[0019] (3) Separation: filtering the enzymatic hydrolyzate and centrifuging to obtain soybean peptide flavor liquid;

[0020] (4) Step-by-step preparation: mix the soybean peptide flavor liquid, maltodextrin and brown sugar syrup, heat at 85°C to 110°C for 15 to 30 minutes, add stevioside, edible salt and monosodium glutamate, and heat at 75°C to 105°C for 20 to 40 minutes;

[0021] (5) Mild reaction: cool to 71°C to 87°C, add flavoring, potassium sorbate and DL-malic acid, stir evenly, and react gently for 30 to 55 minutes;

[0022] (6) Sterilization: 82℃~103℃ for 30~50min;

[0023] (7) Cooling: Cool the material to 42°C to 56°C within 7 to 15 minutes;

[0024] (8) Filling.

[0025] In the method for preparing the food-derived placebo for the above-mentioned clinical trial of stroke-refreshing solution:

[0026] Preferably, in step (2), the enzyme is inactivated at 85-100° C. for 15-40 min.

[0027] Preferably, in step (3), 200-500 mesh filter cloth is used for filtration, and during centrifugal separation, the centrifuge speed is 4500-10000 rpm.

[0028] Preferably, in steps (1) to (2), an enzymatic hydrolysis device having heating and cooling functions and a stirrer is used, such as an enzymatic hydrolysis tank having heating and cooling functions and a stirrer.

[0029] Preferably, in steps (4) to (5), a temperature-controllable batching device equipped with a stirrer is used, such as a temperature-controllable reaction tank equipped with a stirrer.

[0030] Preferably, in steps (6) to (7), a sterilization device having heating and cooling functions and a stirrer is used, such as a sterilization tank having heating and cooling functions and a stirrer.

[0031] Preferably, in step (8), filling and dispensing equipment with quantitative weighing is used for filling.

[0032] The present invention has the following beneficial effects: the inventor of the present application has prepared a food-based placebo through a large number of experiments, using a certain ratio and process. The prepared placebo is similar to the experimental drug stroke awakening solution in appearance, smell, taste, etc., and is not easily identified by researchers and subjects. It can prevent bias and unblinding in double-blind, placebo-controlled clinical trials to the greatest extent, thereby ensuring the quality of clinical trials. It is a creation in the preparation of traditional Chinese medicine placebos. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is described in detail below in conjunction with specific embodiments, so that those skilled in the art can better understand and implement the technical solution of the present invention. The reagents or materials used in the embodiments, unless otherwise specified, are all from commercial channels.

[0034] Example 1

[0035] The food-derived placebo for the clinical trial of the stroke-refreshing solution provided in this embodiment is prepared from the following raw materials in percentage by weight:

[0036]

[0037]

[0038] The protease used was commercially available papain.

[0039] The edible flavor adopts commercially available ginseng flavor.

[0040] The method for preparing the food-derived placebo for the clinical trial of the stroke refreshing solution comprises the following steps:

[0041] (1) Pretreatment: Soy protein powder and gluten powder were mixed evenly according to the proportion and stored at 55°C for 60 min;

[0042] (2) Enzymatic hydrolysis: mix soy protein powder, protease and water, stir, perform enzymatic hydrolysis at 55°C for 4 h, and inactivate the enzyme at 90°C for 35 min;

[0043] (3) Separation: The enzymatic hydrolysate is separated using a 400-mesh filter cloth at 6000 rpm to obtain a soybean peptide flavor liquid;

[0044] (4) Step-by-step preparation: mix soybean peptide flavor liquid, maltodextrin, and brown sugar syrup, heat at 110°C for 20 min, add stevioside, edible salt, and monosodium glutamate, and heat at 75°C for 40 min;

[0045] (5) Mild reaction: cool to 75°C, add flavoring, potassium sorbate, and DL-malic acid, stir evenly, and react gently for 55 minutes;

[0046] (6) Sterilization: 85°C for 50 min;

[0047] (7) Cooling: Cool the material to 50°C within 10 minutes;

[0048] (8) Filling.

[0049] In steps (1) to (2), an enzymolysis device with heating and cooling functions and a stirrer is used, such as an enzymolysis tank with heating and cooling functions and a stirrer.

[0050] In steps (4) to (5), a temperature-controllable batching device with a stirrer is used, such as a temperature-controllable reaction tank with a stirrer.

[0051] In steps (6) to (7), a sterilization device with heating and cooling functions and a stirrer is used, such as a sterilization tank with heating and cooling functions and a stirrer.

[0052] In step (8), filling and dispensing equipment with quantitative weighing is used for filling.

[0053] Example 2

[0054] The food-derived placebo for the clinical trial of the stroke-refreshing solution provided in this embodiment is prepared from the following raw materials in percentage by weight:

[0055]

[0056]

[0057] The protease is trypsin.

[0058] The edible flavor adopts commercially available ginseng flavor.

[0059] The method for preparing the food-derived placebo for the clinical trial of the stroke refreshing solution comprises the following steps:

[0060] (1) Pretreatment: Soy protein powder and gluten powder were mixed evenly according to the proportion and stored at 75°C for 30 min;

[0061] (2) Enzymatic hydrolysis: mix soy protein powder, protease and water, stir, perform enzymatic hydrolysis at 60°C for 8 h, and inactivate the enzyme at 85°C for 40 min;

[0062] (3) Separation: The enzymatic hydrolyzate is separated using a 500-mesh filter cloth at 5500 rpm to obtain a soybean peptide flavor liquid;

[0063] (4) Step-by-step preparation: mix soybean peptide flavor liquid, maltodextrin, and brown sugar syrup, heat at 100°C for 15 min, add stevioside, edible salt, and monosodium glutamate, and heat at 105°C for 20 min;

[0064] (5) Mild reaction: cool to 80°C, add flavoring, potassium sorbate, and DL-malic acid, stir evenly, and react gently for 50 min;

[0065] (6) Sterilization: 103°C for 30 min;

[0066] (7) Cooling: Cool the material to 42°C within 15 min;

[0067] (8) Filling.

[0068] In steps (1) to (2), an enzymolysis tank with heating and cooling functions and a stirrer is used.

[0069] In steps (4) to (5), a temperature-controllable reaction tank equipped with a stirrer is used.

[0070] In steps (6) to (7), a sterilization tank with heating and cooling functions and a stirrer is used.

[0071] In step (8), filling and dispensing equipment with quantitative weighing is used for filling.

[0072] Example 3

[0073] The food-derived placebo for the clinical trial of the stroke-refreshing solution provided in this embodiment is prepared from the following raw materials in percentage by weight:

[0074]

[0075] The proteases are pepsin and trypsin.

[0076] The edible flavor adopts commercially available ginseng flavor.

[0077] The method for preparing the food-derived placebo for the clinical trial of the stroke refreshing solution comprises the following steps:

[0078] (1) Pretreatment: Soy protein powder and gluten powder were mixed evenly according to the proportion and stored at 60°C for 50 min;

[0079] (2) Enzymatic hydrolysis: mix soy protein powder, protease and water, stir, perform enzymatic hydrolysis at 66°C for 6 h, and inactivate the enzyme at 90°C for 30 min;

[0080] (3) Separation: The enzymatic hydrolysate is separated using a 300-mesh filter cloth at 5000 rpm to obtain a soybean peptide flavor liquid;

[0081] (4) Step-by-step preparation: mix soybean peptide flavor liquid, maltodextrin, and brown sugar syrup, heat at 90°C for 30 min, add stevioside, edible salt, and monosodium glutamate, and heat at 100°C for 30 min;

[0082] (5) Mild reaction: cool to 87°C, add flavoring, potassium sorbate, and DL-malic acid, stir evenly, and react gently for 30 min;

[0083] (6) Sterilization: 90°C for 50 min;

[0084] (7) Cooling: Cool the material to 50°C within 10 minutes;

[0085] (8) Filling.

[0086] In steps (1) to (2), an enzymolysis device with heating and cooling functions and a stirrer is used, such as an enzymolysis tank with heating and cooling functions and a stirrer.

[0087] In steps (4) to (5), a temperature-controllable batching device with a stirrer is used, such as a temperature-controllable reaction tank with a stirrer.

[0088] In steps (6) to (7), a sterilization device with heating and cooling functions and a stirrer is used, such as a sterilization tank with heating and cooling functions and a stirrer.

[0089] In step (8), filling and dispensing equipment with quantitative weighing is used for filling.

[0090] Example 4

[0091] In this example, the effect of the placebo was evaluated in the following manner.

[0092] 1. Manual sensory evaluation

[0093] Manual sensory evaluation is to manually describe and score the taste, smell and color of Chinese medicinal materials and placebos. An ideal placebo needs to be completely consistent with the test drug in appearance, color, smell, taste, packaging, usage and dosage. There is currently no recognized evaluation method and standard for the quality evaluation of placebos. It is difficult to prepare an ideal Chinese medicine placebo. The evaluation of Chinese medicine placebos should have its own characteristics. When it is impossible to be completely consistent with the test drug, the placebo should be made as close as possible to the appearance and taste of Chinese medicine in the general sense.

[0094] The specifications and packaging of the placebo involved in this embodiment adopt the same standards as the clinical trial drugs, so the evaluation of the placebo mainly focuses on the appearance, color, taste, smell and whether it is easy to be detected.

[0095] The effect evaluation index of the placebo includes two parts: consistency evaluation and simulation test actual situation. In this example, the test drug and the placebo are first blinded, and multiple evaluators are invited to simultaneously evaluate the appearance and smell, sweetness, bitterness, and comprehensive taste of the blinded test drug and placebo.

[0096] (1) Placebo and test drug consistency test

[0097] The researchers blinded the experimental drug (Stroke Brain Refresher developed by the Second Affiliated Hospital of Guangzhou University of Chinese Medicine) and the placebo (Example 2), and the evaluators evaluated the blinded experimental drug and placebo, including the appearance and taste of the liquid. The results are shown in Table 1-2 below:

[0098] Table 1 Evaluation of the consistency of appearance between placebo and test drug

[0099]

[0100] Table 2 Evaluation of the taste consistency between placebo and test drug

[0101]

[0102] Note: There are 10 people participating in the tasting, and the data in the table represents how many people voted.

[0103] (2) Actual situation of simulation test

[0104] Prepare 14 transparent plastic cups, 7 of which contain test drugs and 7 contain placebos. Use a syringe to take a small amount of test drugs or placebos and inject them into the cups. The subjects are asked to take one of the cups and taste it, and then judge whether it is test drugs or placebos. The results in Table 3 show that only 25.0% of the subjects can correctly distinguish between test drugs and placebos, and 75.0% of the subjects cannot distinguish or make incorrect distinctions.

[0105] Table 3 Test results of the actual situation of the simulation test

[0106]

[0107] Note: Fisher's Exact Test, P=0.001

[0108] According to the test of the reviewers, the color and clarity of the placebo and the test drug are relatively consistent. In terms of taste, the sweetness, smell and overall taste are not consistent, and the bitterness is relatively consistent. The placebo tastes sweeter, the test drug tastes bitterer, and the test drug has a stronger aroma. When the reviewers taste the test drug or the placebo independently, the reviewers who try the test drug can basically confirm that it is the test drug, and the reviewers who try the placebo can basically not distinguish it from the placebo.

[0109] 2. Quality Index Evaluation

[0110] According to the standards of "Compound Seasoning GB 31644" and "National Food Safety Standard Food Microbiology Test Colony Count Determination GB 4689.2-2016", some quality indicators of the placebo of the present invention were tested, and the results were in compliance with the regulations. The results are shown in Table 4.

[0111] Table 4 Placebo product inspection report

[0112] Inspection items Require Inspection basis Test results Single evaluation Color Meet the standard requirements GB 31644 Comply with standards qualified Taste, smell Meet the standard requirements GB 31644 Comply with standards qualified state Meet the standard requirements GB 31644 Comply with standards qualified Total colony count CFU / g ≤30000 GB 4789.2-2016 ＜10 qualified Coliform group MPN / g ≤0.3 GB 4789.2-2016 ＜0.3 qualified

[0113] The above embodiments are only used to illustrate the present invention, and the protection scope of the present invention is not limited to the above embodiments. A person skilled in the art can achieve the purpose of the present invention based on the above disclosure of the present invention, and any improvement and deformation based on the concept of the present invention shall fall within the protection scope of the present invention, and the specific protection scope shall be subject to the claims.

Claims

1. A food-derived placebo for clinical trials of stroke-refreshing liquid, characterized by: The placebo is mainly made of the following raw materials in percentage by weight: 2-15% soy protein powder, 2-9% gluten powder, 0.04-0.3% protease, 4-12% maltodextrin, 2-9% brown sugar syrup, 0.2-1.4% stevioside, 0.05-0.1% edible salt, 0.01-0.3% monosodium glutamate, 1-8% edible flavor, 0.01-0.3% potassium sorbate, 0.01-0.8% DL-malic acid, and the balance is made up of water; The protease is one or more of papain, pepsin and trypsin; The essence is ginseng essence; The food-derived placebo for the clinical trial of stroke-refreshing solution is prepared by the following method: (1) Pretreatment: Mix the soybean protein powder and gluten powder in a certain proportion to obtain a mixed soybean protein powder, and store the mixed soybean protein powder at 55 to 75° C. for 30 to 60 minutes; (2) Enzymatic hydrolysis: Mix the mixed soybean protein powder, protease and water, stir, and perform enzymatic hydrolysis at 50°C to 66°C for 3 to 8 hours, and then inactivate the enzyme; (3) Separation: filtering the enzymatic hydrolyzate and centrifuging to obtain soybean peptide flavor liquid; (4) Step-by-step preparation: mix the soybean peptide flavor liquid, maltodextrin and brown sugar syrup, heat at 85°C to 110°C for 15 to 30 minutes, add stevioside, edible salt and monosodium glutamate, and heat at 75°C to 105°C for 20 to 40 minutes; (5) Mild reaction: cool to 71°C to 87°C, add flavoring, potassium sorbate and DL-malic acid, stir evenly, and react gently for 30 to 55 minutes; (6) Sterilization: 82℃~103℃ for 30~50min; (7) Cooling: Cool the material to 42°C to 56°C within 7 to 15 minutes; (8) Filling.

2. According to claim 1, the food-derived placebo for clinical trial of stroke-refreshing solution is characterized by: The mass ratio of the soy protein powder to the gluten powder is 3-6:

1.

3. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 1 or 2, characterized in that: The following steps are involved: (1) Pretreatment: Mix the soybean protein powder and gluten powder in a certain proportion to obtain a mixed soybean protein powder, and store the mixed soybean protein powder at 55 to 75° C. for 30 to 60 minutes; (2) Enzymatic hydrolysis: Mix the mixed soybean protein powder, protease and water, stir, and perform enzymatic hydrolysis at 50°C to 66°C for 3 to 8 hours, then inactivate the enzyme; (3) Separation: filtering the enzymatic hydrolyzate and centrifuging to obtain soybean peptide flavor liquid; (4) Step-by-step preparation: mix the soybean peptide flavor liquid, maltodextrin and brown sugar syrup, heat at 85°C to 110°C for 15 to 30 minutes, add stevioside, edible salt and monosodium glutamate, and heat at 75°C to 105°C for 20 to 40 minutes; (5) Mild reaction: cool to 71°C to 87°C, add flavoring, potassium sorbate and DL-malic acid, stir evenly, and react gently for 30 to 55 minutes; (6) Sterilization: 82℃~103℃ for 30~50min; (7) Cooling: Cool the material to 42°C to 56°C within 7 to 15 minutes; (8) Filling.

4. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: When the enzyme is inactivated in step (2), the enzyme is inactivated at 85 to 100° C. for 15 to 40 minutes.

5. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: In step (3), 200-500 mesh filter cloth is used for filtration, and during centrifugal separation, the centrifuge speed is 4500-10000 rpm.

6. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: In steps (1) to (2), an enzymatic hydrolysis device with heating and cooling functions and a stirrer is used.

7. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: In steps (4) to (5), a batching device with temperature control and a stirrer is used.

8. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: In steps (6) to (7), a sterilization device with heating and cooling functions and a stirrer is used.

9. The method for preparing the food-derived placebo for clinical trial of stroke-refreshing solution according to claim 3, characterized in that: The filling in step (8) adopts a filling and dispensing device with quantitative weighing.

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