Glycoside drug oral preparation for treating cerebral ischemic stroke during recovery period and preparation method therefor

Abstract

A glycoside drug oral preparation for treating cerebral ischemic stroke during the recovery period and a preparation method therefor. The glycoside drug is (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol. The oral preparation can provide a high drug loading capacity, release the drug within a short time period, and ensure rapid drug dissolution, and the prepared tablet has high stability, involves a simple preparation process and is suitable for large-scale industrial production.

Description

An oral preparation of glycoside drugs for treating ischemic stroke recovery period and its preparation method Technical Field

[0001] The present invention belongs to the field of medical technology and relates to an oral preparation of a glycoside drug and a preparation method thereof. Background Art

[0002] Stroke, commonly known as a stroke, is an acute cerebrovascular disease caused by a sudden rupture of a cerebral blood vessel or a blockage that prevents blood flow to the brain, leading to brain damage. Stroke is now the leading cause of death in China. According to statistics, over 2 million people die from stroke in China each year. Stroke is characterized by high morbidity, mortality, and disability, placing a significant burden on families and society. Stroke can be categorized by pathology: ischemic stroke and hemorrhagic stroke. Ischemic stroke has a higher incidence than hemorrhagic stroke and accounts for approximately 60% to 70% of all strokes.

[0003] Ischemic stroke is clinically classified into the ultra-early, acute, recovery, and sequelae stages. The ultra-early stage is within 6 hours of onset; the acute stage is between 6 hours and 2 weeks of onset; the recovery stage is between 2 weeks and 6 months of onset; and the sequelae stage is beyond 6 months. Currently, the main treatment options for patients in the recovery stage of ischemic stroke are rehabilitation therapy and secondary prevention of stroke. The main therapeutic drugs used are butylphthalide capsules, clopidogrel tablets, and Chinese patent medicines such as danshen and ginkgo. Currently, there is a shortage of therapeutic drugs for ischemic stroke, and there is an urgent need to increase scientific research efforts to develop new drugs with therapeutic effects and fewer side effects. Research and development of drugs for the treatment of stroke is active both domestically and internationally, and there is a high demand for these drugs in clinical practice.

[0004] Glycoside compound (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol, the compound structure is as follows:

[0005] Research on this compound has been extensively conducted. For example, the previously applied Chinese patent CN109988201B discloses and authorizes the glycoside compound; Chinese patent CN112625073B discloses and authorizes the preparation method of the glycoside compound; Chinese patent CN109988200B discloses and authorizes the use of the glycoside compound for preventing and treating ischemic cerebrovascular diseases, especially stroke; Chinese patent CN109988202B discloses and authorizes the use of the glycoside compound for preventing and treating lower limb ischemic microcirculatory disorders, especially diabetic foot gangrene; and Chinese patent CN113456654B discloses and authorizes the injectable dosage form of the glycoside compound.

[0006] For ischemic stroke, our research found that this glycoside compound can achieve neuroprotection by inhibiting the excitotoxicity and oxidative stress damage of nerve cells caused by hypoxia and regulating the expression of cellular inflammatory factors and neurotrophic factors, thereby achieving the effect of treating ischemic stroke.

[0007] Currently, the injectable dosage form of this glycoside compound has entered Phase II clinical research and is used to treat patients in the acute stage of ischemic stroke.

[0008] For patients in the recovery period of ischemic stroke, the use of injections is extremely inconvenient. Therefore, developing an oral medication for the treatment of ischemic stroke in the recovery period has great clinical value. However, when glycoside compounds are prepared as oral preparations, there are problems with the drug's loading capacity, dissolution, and stability. How to prepare these glycoside compounds into an oral preparation that is easy to use and easy to carry is a problem that needs to be solved urgently.

[0009] Summary of the Invention

[0010] The main purpose of the present invention is to provide an oral glycoside preparation and a preparation method thereof which is easy to take, has a large drug loading capacity, fast dissolution, good stability, a simple preparation process and is suitable for large-scale industrial production.

[0011] In a first aspect, the present invention provides an oral preparation of a glycoside drug, wherein the preparation comprises a glycoside drug active ingredient, a filler, a binder, a disintegrant and a lubricant.

[0012] The active ingredient of the glycoside drug is the glycoside compound (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol, with the structural formula as follows

[0013] The filler is selected from one or more of lactose, starch and dextrin, more preferably lactose and / or directly compressible starch; the contents of the filler and / or directly compressible starch in the oral preparation are 6-25 wt% and 0-20 wt%, respectively.

[0014] The binder is hydroxypropyl cellulose, and its content in the oral preparation is 0 to 8% by weight.

[0015] The disintegrant is selected from povidone, and its content in the oral preparation is 0-8% by weight.

[0016] The lubricant is magnesium stearate, and its content in the oral preparation is 0.1-2% by weight.

[0017] The content of the active ingredient glycoside compound in the oral preparation is 40 to 90 weight %, preferably 50 to 80 weight %.

[0018] Each unit of the oral preparation contains 100 mg to 600 mg of the compound of formula (I), preferably 150 mg to 550 mg.

[0019] In a second aspect, the present invention provides a method for preparing the oral preparation according to the first aspect, the method comprising the following steps:

[0020] S01 premixing the active ingredient of the glycoside drug with a filler, a binder and / or a disintegrant at room temperature;

[0021] S02 Pass the premixed mixture from S01 through a 30-mesh sieve;

[0022] S03: Mix the sieved mixture in S02 thoroughly again;

[0023] S04 adds the mixture of S03 to the lubricant for lubrication;

[0024] S05: Mix the lubricated mixture evenly and compress it into tablets using a tablet press.

[0025] In a third aspect, the present invention provides a use of the oral glycoside preparation of the first aspect for preparing a medicament for treating stroke, wherein the stroke is preferably ischemic stroke in the recovery phase.

[0026] Furthermore, the glycoside oral preparation includes one or more of lozenges, effervescent tablets, sustained-release tablets, controlled-release tablets, capsules, enteric-coated tablets, chewable tablets, pills and soft capsules.

[0027] The beneficial effects of the oral glycoside preparation of the present invention are: ① the prepared oral tablets dissolve quickly and have stable quality; ② the glycoside drug loading is high, convenient to take, and meets clinical drug administration needs; ③ the preparation process is simple and can be industrialized on a large scale. DETAILED DESCRIPTION

[0028] The following is a further explanation of the invention contents of the present invention through specific embodiments, but the protection scope of the present invention is not limited to the following examples. According to the inventive ideas and full content of the present invention, the various technical features in the following examples can be appropriately combined / replaced / adjusted / modified, etc., and still fall within the scope of protection of the present invention.

[0029] Example 1

[0030] Preparation process: According to the formula, the active ingredient of the glycoside drug is premixed with lactose, directly compressible starch and hypromellose in sequence at room temperature, passed through a 30-mesh sieve and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 800 mg; the oral tablets are packaged in bottles with a desiccant.

[0031] Example 2

[0032] Preparation process: According to the formula, the active ingredients of the glycoside drug are premixed with lactose and directly compressible starch in sequence at room temperature, sieved and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 400 mg; the oral tablets are packaged in bottles with a desiccant.

[0033] Example 3

[0034] Preparation process: According to the formula, the active ingredients of the glycoside drug are premixed with lactose and directly compressible starch in sequence at room temperature, sieved and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 340 mg; the oral tablets are packaged in bottles with a desiccant.

[0035] Example 4

[0036] Preparation process: According to the formula, the active ingredients of the glycoside drug are premixed with lactose and povidone in sequence at room temperature, sieved and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 680 mg; the oral tablets are packaged in bottles with a desiccant.

[0037] Example 5

[0038] Preparation process: According to the formula, the active ingredients of the glycoside drug are premixed with lactose, directly compressible starch and povidone in sequence at room temperature, sieved and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 340 mg; the oral tablets are packaged in bottles with a desiccant.

[0039] Example 6

[0040] Preparation process: According to the formula, the active ingredients of the glycoside drug are premixed with lactose and directly compressible starch in sequence at room temperature, sieved and mixed until uniform, magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to obtain white glycoside oral tablets with a tablet weight of 680 mg; the oral tablets are packaged in bottles with a desiccant.

[0041] Example 7

[0042] Preparation process: According to the formula, the glycoside active ingredient is premixed with lactose and directly compressible starch at room temperature, sieved, and mixed until uniform. Magnesium stearate is added for lubrication and mixing, and then tableted using a tablet press to produce off-white glycoside oral tablets weighing 340 mg. The tablets are then coated with the Opadry film coating premix at a coating weight gain of 2-5%. The tablets are packaged in bottles with a desiccant.

[0043] Example 8 Dissolution Experiment

[0044] The oral tablet samples prepared in Examples 1 to 7 were tested according to the dissolution assay method (Chinese Pharmacopoeia 2020 General Rules 0931 Second Method), and the results are as follows:

[0045] The results show that the dissolution rates of the glycoside oral tablets of each embodiment of the present invention are good and can be rapidly dissolved in a short time.

[0046] Example 9 Accelerated stability test

[0047] The oral tablet samples prepared in Examples 1 to 7 were placed at (40°C, RH 75%) and subjected to accelerated stability testing. The results are as follows:

[0048] The results showed that, under the conditions of accelerated stability study, the content and total impurities of the glycoside oral tablets of Examples 4 and 5 containing the disintegrant povidone did not change significantly, but the tablet surface appearance lost its gloss. The formulations of Examples 1-3, as well as the formulations of Examples 6-7, showed no obvious degradation and maintained stable quality.

[0049] Example 10: Drug efficacy experiment

[0050] 1. Experimental Materials

[0051] Drug preparation: According to the preparation process of Example 1-7, prepare oral tablets of glycoside compounds of prescription 1-7 (drug loading 13.5 mg / tablet); positive control drug butylphthalide soft capsule (NBP), specification: 0.1 g / tablet, manufacturer: Shijiazhuang Pharmaceutical Group Enbi Pharmaceutical Co., Ltd.

[0052] Experimental animals: 100 SD rats, SPF grade, male, about 300-330 g, 8-10 weeks old, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0053] Housing conditions: Males and females were housed in separate cages, with no more than five animals per cage. Animal room environmental parameters were recorded during the entire period of housing. Housing environment: Temperature: 20–26°C; Relative humidity: 40%–70%; Ventilation: ≥15 air changes per hour; Feed: Cobalt-60 irradiated sterilized SPF mouse and rat maintenance feed, purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd. Drinking water: Homemade deionized water available ad libitum. Bedding: Corncob bedding, purchased from Dezhou Gumei Agricultural Technology Co., Ltd.

[0054] 2. Animal Model Production

[0055] For the efficacy experiment, male SD rats weighing 280-320 g were used. They were anesthetized with isoflurane and fixed on a surgical board with their backs facing up. The rectal temperature was maintained at 36-37°C with a heating pad. The hair on the neck was trimmed and the rats were disinfected with iodine. The skin was cut open along the midline of the neck, with an incision of approximately 1.5 cm. The subcutaneous tissue was separated with blunt forceps to avoid bleeding. The left common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA) were exposed. The proximal and distal ends of the common carotid artery were ligated, and a slipknot was tied at the distal end (below the bifurcation of the ICA and ECA), approximately 1 cm apart. The external carotid artery was then ligated. A small incision was made in the CCA, and a suture was inserted. The slipknot at the distal end was loosened, and the suture was slowly advanced from the internal carotid artery to the anterior end of the middle cerebral artery. When significant resistance was felt, the advancement was stopped, the suture was secured, and the suture was cut short outside the vessel. After disinfection, the wound was sutured. After 90 minutes, the wound was opened, the suture was removed until it was exposed, a black mark was placed on the suture, and the wound was sutured. Body weights of the rats were recorded on days 1, 4, 7, 10, and 14 after surgery.

[0056] 3. Dosage regimen

[0057] In a rat tMCAO model, SHPL-49 formulations 1-7 and NBP were administered orally at a dose of 90 mg / kg 30 minutes after middle cerebral artery occlusion and reperfusion. The efficacy of administration was monitored for 14 days. Neurological function scores were assessed using the mNSS (mean neural network scale) on days 1, 4, 7, 10, and 14. Rotarod and dark-shuttle tests were performed on day 14. After 14 days of administration, the rats were sacrificed, and infarct volumes were calculated using TTC staining to determine the therapeutic effects of SHPL-49 API, different formulations of SHPL-49, and NBP on ischemic stroke.

[0058] This experiment required 100 male rats, and set up 10 groups: Sham group (sham operation), Model group (model group), the prescriptions of Examples 1-7 and NBP.

[0059] The contents of the butylphthalide soft capsule (NBP) were withdrawn with a 1ml syringe and diluted with corn oil. The concentration was calculated based on the dose and volume before administration, and 1.5ml was administered orally to each rat. Tablets were administered orally based on the dose and rat body weight before administration, with two tablets administered orally to each rat. NBP was prepared immediately before use, and all medications were stored at room temperature, protected from light. The dosage for each rat was 0.30kg. The sham and model groups were each gavaged with 1.5ml of 0.9% sodium chloride injection.

[0060] 4. Observation indicators

[0061] 4.1 Neurological function score

[0062] This study used the modified Neurological Severity Score (mNSS) method to score neurological deficits, with a total score of 18 points. The scoring criteria are detailed in the table below.

[0063] Neurological Injury Severity Score

[0064] This experiment used the mNSS scoring method to comprehensively evaluate the animals' motor, sensory, reflex, and balance functions. Neurological scores were recorded for each group of rats on days 1, 4, 7, 10, and 14 after surgery. Higher scores indicate more severe neurological damage.

[0065] 4.2 Rotarod test

[0066] The rotarod test is a convenient method for assessing motor function in rodents. It assesses an animal's ability to maintain balance on a rotating rod. As the rotation speed increases, the animal's balance becomes more difficult, thus assessing the rodent's motor function.

[0067] Starting from the 11th day of drug administration, rats were trained in a constant speed mode for 3 consecutive days at a training speed of 8 rpm and a training duration of 5 minutes. On the 14th day of drug administration, the constant acceleration mode was used with a test speed of 4-40 rpm and a test duration of 5 minutes. The speed of the rotarod at the time of falling and the time the rat stayed on the rotarod were recorded.

[0068] 4.3 Measurement of cerebral infarction volume

[0069] For acute phase studies, 2,3,5-triphenyltetrazolium chloride (TTC) staining is the preferred method for marking infarcted tissue. TTC works by altering adenosinetriphosphate (ATP) metabolism and is suitable for the early stages of infarction.

[0070] Fourteen days after administration, rats were anesthetized with isoflurane and their brains were quickly removed and frozen at -20°C for 30 minutes. The olfactory bulb, cerebellum, and brainstem were removed and average coronal sections were cut into six slices approximately 2 mm thick. The sections were then quickly placed in 2% tetrazolium chloride (TTC) staining solution for staining. The sections were incubated at 37°C in the dark for 7 minutes, flipped, and incubated for another 7 minutes. The TTC solution was discarded, and an appropriate amount of 4% paraformaldehyde was added. The sections were fixed for 1 hour and then observed. Because the ischemic portion of the brain sections is not stained, while the non-ischemic portion appears red, TTC-stained brain sections were photographed using a digital camera. Adobe Photoshop software was used to calculate the ischemic area in the rat brain sections, and the infarct volume was calculated according to the following formula.

[0071] Where h represents the thickness of each brain slice, Sn and Sn+1 represent the infarct area of ​​adjacent brain slices, respectively.

[0072] 4.4 Statistical analysis

[0073] GraphPad Prism 9.0 software was used for post-experimental statistical analysis of all evaluation indicators. Neurological scores and the number of errors in the dark-shuttle test are expressed as median ± interquartile range (IQR). Infarct volume, rat body weight, latency in the dark-shuttle test, and rotarod test are expressed as mean ± standard deviation (SD). Statistical differences were analyzed using one-way analysis of variance (ANOVA) and two-way ANOVA. *** P<0.001, # P<0.05, ## P<0.01, ### P<0.001 indicates a significant difference. *Compared with the control group (sham group), # Compared with the model group. ns: P>0.05, indicating no significant difference.

[0074] 5. Experimental Results

[0075] 5.1 Neurological function score

[0076] Summary of mNSS neurological score results after 14 consecutive days of drug administration Note: Two-way ANOVA statistical analysis. *** P < 0.001, Model vs Sham; # P<0.05, ## P < 0.01, ### P < 0.001 for Prescriptions 1-7 and NBP vs. Model. P > 0.05 for all other comparisons. N: Number of animals treated for 14 consecutive days.

[0077] According to the table above, compared with the Sham group, the neurological score of the Model group administered with 0.9% sodium chloride by gavage was significantly increased, indicating that the tMCAO model was successfully established ( *** P < 0.001). On days 7, 10, and 14 of administration, oral administration of 90 mg / kg of prescriptions 1-7 and NBP significantly reduced neurological scores and improved neurological deficits compared with the Model group. Prescriptions 3, 4, 6, and 7 were superior to NBP in improving neuromotor behavior in tMCAO rats.

[0078] Based on the above data analysis, prescription 1-7 oral tablets at a dose of 90 mg / kg for 14 days can significantly improve the neuromotor behavior of tMCAO and have a therapeutic effect on cerebral ischemia.

[0079] 5.2 Rotarod test results

[0080] Summary of Rotarod test results after 14 days of continuous drug administration Note: Two-way ANOVA statistical analysis. *** P < 0.001, Model vs Sham; # P<0.05, ## P < 0.01 for Prescriptions 1-7 and NBP vs. Model. P > 0.05 for all other comparisons. N: Number of animals treated for 14 consecutive days.

[0081] Compared with the Sham group, the rats in the Model group spent significantly less time on the rod and their speed of rotating the rod when falling was significantly reduced ( ***P<0.001); Compared with the Model group, the time that rats in the Prescription 1-7 groups stayed on the rod was significantly prolonged, and the speed of the rotating rod when rats in the Prescription 1-7 and NBP groups fell was significantly increased. ( ## P < 0.01, # P<0.05), indicating that prescriptions 1-7 can significantly improve the motor coordination ability of tMCAO rats.

[0082] 5.3 Cerebral infarction volume

[0083] Summary table of infarct volume results after 14 days of continuous administration Note: One-way analysis of variance (ANOVA) statistical method. *** P < 0.001, Model vs Sham; # P<0.05, ## P<0.01, ### P < 0.001 for prescriptions 1-7 and NBP vs Model. P > 0.05 for all others.

[0084] N: The number of animals treated with the drug for 14 consecutive days.

[0085] The results showed that compared with the Sham group, the cerebral infarction volume of rats in the Model group was significantly increased ( *** P<0.001), indicating that the tMCAO model was successfully established. Compared with the Model group, oral administration of 90 mg / kg of prescriptions 1-7 and NBP to rats significantly reduced the infarct volume of tMCAO rats ( # P<0.05, ## P<0.01, ### P < 0.001), and prescriptions 1-7 were superior to NBP, indicating that prescriptions 1-7 at 90 mg / kg could significantly reduce the infarct volume in tMCAO rats.

[0086] The above data analysis shows that prescription 1-7 oral tablets can significantly reduce the infarct volume after cerebral ischemia injury in rats, improve motor coordination ability, have obvious neuroprotective function in rats with cerebral ischemia injury, and have a therapeutic effect on cerebral ischemia.

Claims

1. An oral preparation of glycosides for treating the recovery period of ischemic stroke, characterized in that: The active ingredient of the glycoside drug is the glycoside compound SHPL-49, such as (2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol represented by the general formula (I), and the content of the active ingredient glycoside compound in the oral preparation is 40 to 90 weight percent.

2. The oral glycoside preparation according to claim 1, comprising a filler, a binder, a disintegrant and a lubricant.

3. The oral glycoside preparation according to claim 2, wherein the filler is selected from one or more of lactose, starch and dextrin, more preferably lactose and / or directly compressible starch; the content of the filler and / or directly compressible starch in the oral preparation is 6-25 wt% and 0-20 wt%, respectively.

4. The glycoside oral preparation according to any one of claims 2 to 3, wherein the binder is hydroxypropyl cellulose, and its content in the oral preparation is 0 to 8% by weight.

5. The glycoside oral preparation according to any one of claims 2 to 4, wherein the disintegrant is selected from povidone, and its content in the oral preparation is 0 to 8% by weight.

6. The glycoside oral preparation according to any one of claims 2 to 5, wherein the lubricant is magnesium stearate, and its content in the oral preparation is 0.1-2% by weight.

7. A method for preparing the oral preparation according to claim 6, comprising the following steps: S01 premixing the active ingredient of the glycoside drug with a filler, a binder and / or a disintegrant at room temperature; S02 Pass the premixed mixture from S01 through a 30-mesh sieve; S03: Mix the sieved mixture in S02 thoroughly again; S04 adds the mixture of S03 to the lubricant for lubrication; S05: Mix the lubricated mixture evenly and compress it into tablets using a tablet press.

8. Use of the glycoside oral preparation according to claim 6 in the preparation of a medicament for treating cerebral stroke.

9. The use according to claim 8, wherein the stroke is ischemic stroke in the recovery period.

10. The use according to claim 8 or 9, wherein the glycoside oral preparation comprises tablets, capsules, and granules. Tablets include but are not limited to orally disintegrating tablets, sublingual tablets, dispersible tablets, effervescent tablets, sustained-release tablets, controlled-release tablets, and enteric-coated tablets; capsules include but are not limited to hard capsules, sustained-release capsules, controlled-release capsules, and enteric-coated capsules; and granules include but are not limited to one or more of soluble granules, effervescent granules, enteric-coated granules, and sustained-release granules.

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