CN114010575A

CN114010575A - Novel formulations of neostigmine and derivatives thereof

Info

Publication number: CN114010575A
Application number: CN202111230665.8A
Authority: CN
Inventors: 张译芳; 江华; 胡华胜; 王蓉玲
Original assignee: Jiangsu Tieqi Pharmaceutical Technology Co ltd
Current assignee: Jiangsu Tieqi Pharmaceutical Technology Co ltd
Priority date: 2021-10-22
Filing date: 2021-10-22
Publication date: 2022-02-08

Abstract

The invention provides a novel preparation of neostigmine and derivatives thereof. The preparation comprises sublingual mucosa preparation, nasal mucosa preparation, rectal mucosa preparation, vaginal and uterine mucosa preparation and buccal mucosa preparation which take neostigmine or neostigmine derivatives as active ingredients. The preparation is administrated through any mucous membrane route of buccal mucosa, sublingual mucosa, nasal mucosa, rectal mucosa or vagina and uterine mucosa, and has great clinical application value in preparing medicines for treating myasthenia gravis, postoperative abdominal distension and urinary retention, supraventricular paroxysmal tachycardia treatment and detoxification when tubocurarine is excessive.

Description

Novel formulations of neostigmine and derivatives thereof

Technical Field

The invention relates to the technical field of pharmaceutical preparations, in particular to a novel preparation of neostigmine and derivatives thereof and application thereof.

Background

Neostigmine (Neostigmine) has reversible cholinesterase inhibition effect, so that acetylcholine is not subjected to enzymolysis, exists in cholinergic nerve endings for a long time, and has the effect of exciting smooth muscle; the N2 choline receptor on the motor end plate of skeletal muscle can be directly excited, so that the effect on the skeletal muscle is stronger, and the miosis force is smaller; it is used for myasthenia gravis, postoperative abdominal distention and urinary retention, and for the treatment of supraventricular paroxysmal tachycardia and excessive Toxodrine. Neostigmine bromide has the following structural formula:

clinical administration routes of drugs are mainly divided into gastrointestinal administration and parenteral administration, such as oral administration, injection administration, transdermal administration, inhalation administration and the like. The oral administration is convenient to carry and use, and does not cause physiological discomfort to patients, so that the oral administration has higher patient compliance, but after the oral administration, the medicine is dissolved into blood in the gastrointestinal tract, and finally, the target effect taking process is achieved, the effect taking is slow, so that the oral administration is not an optimal administration way for some insoluble medicines or acute diseases, and secondly, because the gastrointestinal tract environment is more complex, and some medicines which are not acid and alkali resistant can generate obvious degradation phenomenon or obvious first pass effect after entering the gastrointestinal tract, the bioavailability is lower. Injection administration is fast acting and accurate in dosage, but has poor patient compliance due to the significant physiological pain it can cause to patients with invasive administration. Therefore, for some drugs with poor oral bioavailability, the development of a non-invasive parenteral administration mode has a very significant clinical value, oral mucosa administration is one of the non-invasive parenteral administration routes, and with the rapid development of novel polymers and drug delivery technologies, the attention of more and more medical workers on how to safely and effectively administer drugs through the oral mucosa route to avoid complex gastrointestinal environment is paid. The clinical application of neostigmine bromide is mainly injection and oral tablets, the injection administration is inconvenient, physiological discomfort can be brought to patients during administration, the medicine cannot be well used for treating diseases, and the oral tablets are difficult to absorb and irregular, so that the noninvasive parenteral administration route has great market potential.

Publication No. CN102258492A discloses a prescription of neostigmine bromide sustained-release tablets which are taken once a day and can be used for treating myasthenia gravis, functional intestinal flatulence after operation and urinary retention and a preparation process thereof. The neostigmine bromide sustained release tablet can overcome the defects of the common tablets of the existing products, and slowly release relatively stable blood concentration and longer action time. The publication number CN103191059A discloses a prescription of neostigmine bromide multivesicular liposome and a preparation method thereof, and the neostigmine bromide multivesicular liposome prepared by the invention can realize the slow release of the drug and prolong the action time and the drug effect time of the drug. The publication number CN104523567A provides a neostigmine bromide composition freeze-dried tablet and a preparation method thereof, the neostigmine bromide composition freeze-dried tablet only needs two auxiliary materials of starch and cane sugar, a freeze-drying process with two liters of reduction and two liters of temperature is adopted, the tablet formability is better by two temperature reduction and two temperature rise, the dissolution rate of the tablet is improved, and the bioavailability of the tablet is improved. Publication No. CN105853370A provides neostigmine bromide powder and a preparation method thereof, wherein the powder comprises the following components in percentage by weight: neostigmine bromide: excipient 0.5: 99.5, the neostigmine bromide powder has simple preparation process, high bioavailability and low production cost. However, the administration route of the product prepared by the invention is still the gastrointestinal tract, namely, the product is inevitably required to pass through the gastrointestinal tract before the target of action is taken to have a remarkable first-pass effect, and finally the bioavailability is low.

Publication No. CN111557906A discloses a neostigmine mesylate injection and a preparation method thereof. The neostigmine methosulfate injection comprises 0.05 to 0.08 percent of neostigmine methosulfate, 0.7 to 0.9 percent of isoosmotic adjusting agent, 0.095 to 0.25 percent of pH adjusting agent and the balance of water for injection; the neostigmine mesylate injection prepared by selecting specific auxiliary materials and reasonably controlling process steps has the advantages of isotonic product, high sterility guarantee level, safety, effectiveness, controllable quality and F₀More than 15, and the sterility guarantee level SAL is less than or equal to 10⁶The prepared injection has low impurity level content, the osmotic pressure ratio of the product is 0.9-1.1, the injection still has high stability after being accelerated for 6 months at 40 ℃, and all impurities are not obviously increased and are not generated newly. Publication No. CN113230206A discloses a neostigmine mesylate composition and a preparation method thereof, comprising: an effective amount of neostigmine mesylate; a chelating agent; a pH adjusting agent; and water. The composition is simple, the using amount is small, the production cost is reduced, the production risk of microorganism and bacterial endotoxin pollution is reduced, the high temperature resistance of the neostigmine mesylate composition is obviously improved, and the amount of the impurity A of the composition after high-temperature sterilization can be reduced by half. Publication No. CN110585143A relates to neostigmine mesylate freeze-dried preparation for injection and a preparation method thereof. Neostigmine methosulfate injection is unstable to light and heat, and mainly degrades 3-hydroxytrimethylidine methosulfate which is an impurity and has a genotoxicity warning structure. In order to improve the stability of the freeze-dried preparation, the inventor carries out dosage form improvement on the freeze-dried preparation, and obtains the freeze-dried preparation product through prescription screening and process improvement. The obtained product has good appearance, good re-solubility and good clarity, the main degradation impurity 3-hydroxytrimethylidene methosulfate is greatly reduced, and the safety and the stability of the product are improved. However, the invention improves the existing preparation on the aspect of improving the stability of neostigmine and analogues thereof, and the prepared product is administrated by injection, can effectively avoid the remarkable first-pass effect of neostigmine and analogues thereof, but is not yet understoodThe problem of clinical compliance caused by invasive administration is solved.

In view of the above, the present invention provides a novel non-invasive parenteral dosage form of neostigmine and its derivatives, and methods of administration and use thereof.

Disclosure of Invention

The invention aims to provide a novel preparation of neostigmine and derivatives thereof and application thereof, the novel preparation for non-invasive parenteral administration has the bioavailability equivalent to that of a neostigmine bromide injection, avoids physiological discomfort brought to patients due to injection administration, improves the medication compliance of the patients, and has great clinical application value.

In order to achieve the above object, the present invention provides a novel formulation of neostigmine and its derivatives. The novel preparation is a sublingual tablet or sublingual membrane, which takes neostigmine or neostigmine derivatives as active ingredients and also comprises a pH regulator and pharmaceutically acceptable auxiliary materials; in a single dose of the novel formulation, the active ingredient is present in an amount of 0.1mg to 20 mg.

As a further improvement of the invention, in the sublingual tablet, the active ingredient is in an amount of 0.1-20 mg, and preferably in an amount of 1-15 mg, and more preferably in an amount of 1-5 mg.

As a further improvement of the invention, the pH regulator is used in an amount of 5% to 15% (w/w), and preferably in an amount of 7% to 13% (w/w), and more preferably in an amount of 9% to 11% (w/w).

As a further improvement of the invention, the acidity of the sublingual tablet is 6-8; the pH regulator is sodium carbonate.

As a further improvement of the invention, in the sublingual tablet, the auxiliary materials comprise 10-90 w/w% of filling agent, 1-10 w/w% of disintegrating agent, 20-60 w/w% of acid-base effervescent agent and 0.5-2 w/w% of lubricating agent, and the sum of all the components is 100% (w/w)

As a further improvement of the invention, the amount of the active ingredient in the sublingual film is 0.1-20 mg, and preferably 1-15 mg, and more preferably 1-5 mg.

As a further improvement of the invention, the acidity of the sublingual membrane is 6-8; the pH regulator is sodium carbonate.

As a further improvement of the invention, in the sublingual membrane, the auxiliary materials comprise 10.0-95.0 w/w% of water-soluble membrane forming materials, 5.0-30.0 w/w% of plasticizers, 0.05-1.5 w/w% of surfactants and a proper amount of stabilizers, flavoring agents and opacifiers, and the sum of the components is 100% (w/w).

As a further improvement of the invention, the active ingredient includes, but is not limited to, one or more combinations of neostigmine, neostigmine bromide, neostigmine methosulfate, and pirstigmine bromide.

In order to realize the purpose, the invention also provides application of neostigmine and a mucous membrane preparation of derivatives thereof. The mucosa preparation takes neostigmine or neostigmine derivatives as active ingredients; the application of the compound in the preparation of the medicine for treating myasthenia gravis, postoperative abdominal distension and urinary retention, supraventricular paroxysmal tachycardia and detoxifying when tubocurarine is excessive through any one mucosal route of oral buccal mucosa, sublingual mucosa, nasal mucosa, rectal mucosa or vaginal and uterine mucosa.

As a further improvement of the invention, the mucosal preparation comprises a novel preparation (sublingual tablet or sublingual membrane) of neostigmine and derivatives thereof, and a nasal mucosal preparation, a rectal mucosal preparation, a vaginal and uterine mucosal preparation and an oral buccal mucosal administration preparation which take the neostigmine or the neostigmine derivatives as active ingredients.

As a further improvement of the invention, the mucosal preparation comprises a solid preparation, a semisolid preparation and the like containing neostigmine and derivatives thereof.

In order to realize the purpose, the invention also provides a mucosal administration method of neostigmine and derivatives thereof. According to the mucosal administration method, neostigmine or neostigmine derivatives are used as active ingredients, and according to the types of mucosa, a new preparation (a sublingual tablet or a sublingual membrane) of the neostigmine and the derivatives thereof is adopted for mucosal administration, or one preparation of a nasal mucosa preparation, a rectal mucosa preparation, a vaginal and uterine mucosa preparation and a cavity buccal mucosa preparation which take the neostigmine or the neostigmine derivatives as the active ingredients is adopted for mucosal administration.

The invention has the beneficial effects that:

1. compared with the common tablets of neostigmine and derivatives thereof prepared by the prior art, the novel preparation sublingual tablet of neostigmine and derivatives thereof provided by the invention has faster dissolution rate. The absolute bioavailability of the sublingual tablet in a rat body of an experimental animal is 91.76%, while the absolute bioavailability of the common neostigmine bromide tablet prepared by the prior art is 32.69%, which shows that the in vivo bioavailability of the sublingual tablet of neostigmine bromide provided by the invention is far superior to that of the common neostigmine bromide tablet, and meanwhile, the sublingual tablet is quick to dissolve and is quickly absorbed by mucous membranes, the drug effect is quickly exerted, the first pass effect and the damage and degradation of gastrointestinal tracts are avoided, the drug dosage is accurate, and the technical problem of lower bioavailability of the oral neostigmine and derivative oral tablets thereof is solved.

2. The new preparation of neostigmine and derivatives thereof provided by the invention has the in vitro steady state permeation rate of 7.2584 mu g cm^-2·h^-1The preparation containing neostigmine and the derivative thereof provided by the invention has good mucosal permeability.

3. The novel preparation of neostigmine and derivatives thereof provided by the invention has the advantages of convenient operation, good reproducibility, strong controllability and lower cost, and related pharmaceutic adjuvants are easy to purchase, thereby having good social and economic benefits. Compared with the common tablet, the tablet has high utilization rate and bioavailability equivalent to that of an injection, so that the administration dosage is greatly reduced; compared with an injection, the injection adopts a mucous membrane approach, effectively avoids the technical defect that a patient generates physiological discomfort caused by injection administration, and improves the medication compliance of the patient, thereby having great clinical application value.

4. The new preparation of neostigmine and its derivative provided by the invention transfers the active ingredient neostigmine and its derivative through a local mucous membrane path by adopting a fat-soluble channel, wherein the absorption amount of the active ingredient is related to the dissociation degree of the active ingredient, the active ingredient existing in an ion form is basically not absorbed by the fat-soluble channel, and the pH value of a medium can influence the dissociation degree of the active ingredient, thereby influencing the mucous membrane absorption amount of the active ingredient. According to local mucous membrane micro-environments at different mucous membrane positions, the pH regulator is used for regulating and controlling the fat-soluble channel so as to realize high-efficiency transfer and higher bioavailability of active ingredients.

Drawings

FIG. 1 shows the results of the measurement of the dissolution curves of the commercially available neostigmine tablets of comparative example 2 and the neostigmine bromide sublingual tablets prepared in examples 1 to 6 in a phosphate buffer solution at pH 7.4.

FIG. 2 is a graph showing the comparison of the time curves of the sublingual bromoneostigmine tablets prepared in example 1 of the present invention, the commercially available bromoneostigmine tablets of comparative example 2, and the commercially available bromoneostigmine injection of comparative example 1 in rat.

FIG. 3 shows the results of the measurement of the dissolution curves of the commercially available neostigmine tablet of comparative example 2, the neostigmine bromide sublingual tablet prepared in example 1 and the sublingual neostigmine bromide film prepared in example 9 in a phosphate buffer solution at pH 7.4.

Fig. 4 is a comparison of in vitro permeation curves for the formulations prepared in example 1 and example 9.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in detail with reference to the accompanying drawings and specific embodiments.

It should be noted that, in order to avoid obscuring the present invention with unnecessary details, only the structures and/or processing steps closely related to the aspects of the present invention are shown in the drawings, and other details not closely related to the present invention are omitted.

In addition, it is also to be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Examples 1 to 6

The composition of the components of examples 1-6 is shown in table 1 below:

table 1 is a table of ingredients for examples 1 to 6

The preparation steps of the preparation methods of the examples 1 to 6 are basically the same, and the main difference is that the formulas are different, and the preparation methods are as follows:

s1, adding neostigmine bromide, mannitol, sodium carboxymethyl starch, sodium carbonate as a pH regulator, citric acid and sodium bicarbonate according to the formula amount into a three-dimensional motion mixer, setting the mixing frequency to be 10Hz, and mixing for 20 min;

s2, adding the magnesium stearate with the prescription amount into the S1 mixed powder, setting the mixing frequency to be 10Hz, and mixing for 5 min;

s3, tabletting the mixed powder by using a high-speed rotary tablet press equipped with 6mm circular dies to obtain the neostigmine bromide sublingual tablet.

Comparative example 1

As comparative example 1(R1, standard: 15 mg. multidot.mL) a commercially available neostigmine bromide injection was used^-1)。

Comparative example 2

A commercially available neostigmine tablet was used as comparative example 2(R2, specification: 15 mg).

The specific methods for evaluating the properties of examples 1 to 6 and comparative examples 1 to 2 are as follows:

1) dissolution curve: the dissolution curve of the sample prepared by the invention and the commercially available neostigmine bromide tablet in artificial saliva is respectively measured by adopting a 2020 edition Chinese pharmacopoeia 0931 second method (slurry method), the volume of a medium is 500mL, the rotating speed is 50rpm, and the sampling time points are 1min, 3min, 5min, 8min, 10min, 15min and 20min, so that the in-vitro dissolution curve comparison research is carried out on the neostigmine bromide sublingual tablet and the commercially available neostigmine bromide tablet.

2) In vivo pharmacokinetic studies in rats: using neostigmine bromide injection (R1, specification: 15 mg. multidot.mL)^-1) And commercially available neostigmine bromide tablets (R2, specification: 15mg) as reference drug, compare the present invention with a self-made neostigmine bromide sublingual tablet (example 1, T, specification: 15mg) and the pharmacokinetic parameters of the reference drug in rats. The scheme is designed as follows: rats were randomly divided into 3 groups, numbered S1-S24, respectively. Before administration, all rats were anesthetized by intraperitoneal injection of ketamine (30mg/kg), and after anesthesia, 1mL of an injection solution of neostigmine bromide was intravenously injected into each of S1-S8 (R1), neostigmine bromide tablets were intragastrically administered into S9-S16 (R2), and sublingual administration of the sublingual membrane of neostigmine bromide (T) into S17-S24 (S17-S24). After administration, each experimental group was administered for 5min, 15min, 30min, 45min, 1h, 2h, 4h, 6h, 8h, 12h, 24h, 36h, 48h, 72h, 0.3mL of blood was collected from rat fundus, the collected blood sample was placed in a heparin-containing centrifuge tube, plasma was collected after centrifugation for 10min (5000 × g), plasma was collected, 200 μ L of plasma was removed, 20 μ L of internal standard solution was added, and after mixing well, a phemenon solid phase extraction column (str-X-CW 33u, 10mg · mL-1) activated in advance with 0.5mL of 5% methanol glacial acetic acid solution, 0.5mL of methanol solution, 0.5mL of 0.2M disodium hydrogen phosphate solution (pH adjusted to 9 with sodium hydroxide) in this order was injected, first, 1mL of 0.2M disodium hydrogen phosphate solution (pH adjusted to 9 with sodium hydroxide): eluting with methanol (90:10, v/v), discarding the eluate, eluting with 1mL of 5% methanol glacial acetic acid solution, collecting the eluate, blowing the eluate with nitrogen flow (40 deg.C), dissolving the residue with 100 μ L of mobile phase, mixing by vortex, and sampling 20 μ L for analysis. The chromatographic conditions were as follows: a Phenomenon SCX C18 column (150X 4.6mm, 5 μm); setting the column temperature to 35 ℃; isocratic elution, 0.5M sodium dihydrogen phosphate (pH adjusted to 3.5 with phosphoric acid): acetonitrile (45:55, v/v); the flow rate is 1 mL/min^-1(ii) a The detection wavelength was 200 nm. Sample determination and analysis of results were as follows: recording the peak area of neostigmine bromide, calculating the concentration of neostigmine bromide in the sample by using a standard curve respectively, calculating pharmacokinetic parameters according to the atrioventricular model and drawing a medicine chamberCurve line.

3) In vitro dissolution studies: neostigmine bromide sublingual tablets were prepared according to the preparation method provided by the present invention in accordance with the recipes of examples 1 to 6, and the in vitro dissolution profiles of the neostigmine bromide sublingual tablets prepared in examples 1 to 6 and the commercially available neostigmine bromide sublingual tablets in artificial saliva were measured, and the results are shown in table 2 and fig. 1.

Table 2 shows the in vitro dissolution of neostigmine bromide sublingual tablets prepared in examples 1-6 and the commercially available neostigmine bromide tablets of comparative example 2 in artificial saliva

Time (min)

1

3

5

8

10

15

Example 1

94.31±1.12

94.85±0.79

94.84±0.68

94.71±1.04

94.94±1.08

95.50±0.84

Example 2

93.10±1.33

93.23±0.76

93.49±0.79

93.58±0.98

93.58±0.80

94.50±1.05

Example 3

92.00±1.08

93.41±0.87

93.94±0.81

93.74±0.64

93.78±0.60

94.37±0.46

Example 4

91.32±1.72

92.95±1.81

93.11±1.94

92.63±2.03

92.69±2.04

92.79±3.05

Example 5

88.70±2.47

92.04±1.13

92.47±1.15

93.67±1.34

93.17±2.44

91.71±1.32

Example 6

91.90±4.99

96.53±1.72

97.57±1.99

98.68±1.82

99.21±1.99

98.72±1.36

Comparative example 2

35.31±2.11

72.58±3.24

89.98±1.09

94.86±1.24

95.63±2.31

95.24±2.01

As is apparent from the results of Table 2 and FIG. 1, the neostigmine bromide sublingual tablets prepared in examples 1-6 were completely dissolved in artificial saliva within 1min, whereas the neostigmine bromide sublingual tablets prepared in comparative example 2 of the prior art were completely dissolved within 5min, indicating that the neostigmine bromide sublingual tablets provided by the present invention were more rapidly dissolved than the existing neostigmine bromide sublingual tablets of comparative example 2.

Pharmacokinetic experiments: comparative pharmacokinetic differences in rats were observed for neostigmine bromide sublingual tablets (group T) prepared in example 1, commercial neostigmine bromide tablets (group R2) of comparative example 2, and neostigmine bromide injection (group R1) of comparative example 1, and the results are shown in table 3 and fig. 2.

Table 3 key pharmacokinetic parameters of neostigmine bromide in rats

Remarks are as follows: f is the absolute bioavailability of the compound,the calculation formula is AUC_{(0- ∞) non-intravenous route of administration}/AUC_{(0- ∞) intravenous route of administration}×100％。

As can be seen from the results of table 3 and fig. 2, the absolute bioavailability of the sublingual tablet of neostigmine bromide provided by the present invention (example 1) in rats was as high as 91.76%, while the absolute bioavailability of the conventional tablet of neostigmine bromide prepared by the prior art of the comparative document 2 in rats was 32.69%, indicating that the bioavailability of the sublingual tablet of neostigmine bromide provided by the present invention in vivo is far superior to that of the conventional common tablet of neostigmine.

Examples 7 to 9

The composition of the components of examples 7-9 is shown in table 1 below:

table 4 shows the composition tables of examples 7 to 9

The preparation steps of the preparation methods of the examples 7 to 9 are basically the same, and the main difference is that the formulas are different, and the preparation methods are as follows:

dispersing hydroxypropyl methylcellulose with the prescription amount into purified water with the temperature of 90 ℃, stirring and dissolving to obtain a colorless transparent solution, then cooling to room temperature, adding neostigmine bromide, polyethylene glycol, aspartame, disodium edetate, polysorbate, titanium dioxide and sodium carbonate with the prescription amount, and stirring for 5 min; then dispersing for 7min by a high-speed disperser (the rotating speed is set to 2000 rpm); and finally obtaining a white emulsion solution, adding the white emulsion solution into a continuous coating machine for continuous coating after ultrasonic defoaming, and cutting after drying to obtain 15 mg/tablet neostigmine bromide sublingual film agent.

The specific method for evaluating the performance is as follows: the in vitro steady state permeation rate of the self-made bromoneostigmine was evaluated using the Franz model. The characteristics of the pig buccal mucosa such as composition, form and permeability barrier function are the closest to those of a human body, so that an in-vitro permeation research model of the pig buccal mucosa as a neostigmine bromide sublingual tablet is selected, and the specific evaluation process is as follows:

(1) treating the mucous membrane of the pig cheek: carefully taking down an electroshocked pig cheek, washing the pig cheek with PBS (0.68 g of monopotassium phosphate, 29.L mL of 0.L mol. L-1 sodium hydroxide solution and water are added) with the pH value of 7.0, carefully removing submucosal tissues, separating out a mucous membrane layer with the thickness of about 600-800 mu m, and placing the mucous membrane layer in the PBS with the pH value of 7.0 for later use.

(2) Evaluation model: franz diffusion cell (12 channels), drug pool was 2mL artificial saliva, and receiving pool was PBS (10mL) at pH 7.0. The whole device is placed in a 37 ℃ constant temperature water bath kettle with a magnetic stirring function, and a magnetic stirrer is placed in a receiving pool at the rotating speed of 300rpm in order to avoid the formation of concentration gradient in the closed receiving pool.

(3) Sample preparation: taking 12 tablets (specification: 0.2mg) of each prescription, respectively placing in a drug pool, starting the experiment after the system is stable, respectively sampling 1mL (simultaneously supplementing equal-volume isothermal receiving medium) at the end of 1h, 2h, 3h, 4h, 5h, 6h and 7h, filtering with a PVDF needle type filter membrane (0.25 μm), and taking the filtrate as a sample solution; an appropriate amount of neostigmine bromide was precisely weighed, dissolved in PBS (pH7.0), and diluted to a concentration of 0.5 mg/mL^-1The solution of (4) as a control solution.

(4) Chromatographic conditions and analysis: the column was Agilent Eclipse XDB-C18 (4.6X 250mm, 5 μm); performing isocratic elution by using acetonitrile-20 mmol.L-1 potassium dihydrogen phosphate solution (pH value is adjusted to 3.0 by phosphoric acid) (10: 90); the detection wavelength is 220 nm; the column temperature was 35 ℃; the flow rate is 1mL min < -1 >; the injection volume was 10. mu.L. Respectively and precisely measuring the sample solution and the reference solution, analyzing the sample injection according to the verified chromatographic conditions, and calculating the cumulative permeation amount of the medicine in unit area at each time point according to an external standard method.

(5) And (3) data analysis: and (3) drawing by taking the unit area accumulated permeation quantity as a vertical coordinate and time as a horizontal coordinate to obtain a drug accumulated permeation curve, and calculating the steady state permeation rate of the drug according to the following formula: p ═ Δ Q/A Δ t

In the formula: p is the steady state permeation rate, Delta Q is the amount of the drug accumulated to permeate the mucous membrane within Delta t time, and A is the effective area of the drug permeating the mucous membrane. The results are shown in Table 3.

Table 5 examination of the effect of the formulation variables on the in vitro permeation rate of the samples (n-12)

Prescription	1h	2h	3h	4h	5h	6h	7h	P(μg·cm^-2·h^-1)
									Example 7	8.99	15.98	22.01	29.01	35.99	42.06	49.21	7.1243
Example 8	7.96	14.28	22.41	28.92	34.52	42.55	50.01	7.1414
									Example 9	8.06	15.11	21.14	29.21	35.02	44.01	51.21	7.2584

From the results in Table 5, it can be seen that different hypromellose dosages have little effect on the rate of sublingual membrane permeation in vitro. And the in vitro steady state permeation rate of the sublingual film of the example 9 reaches 7.2584 mug cm^-2·h^-1The sublingual film containing neostigmine and derivatives thereof has good mucosal permeability.

Referring to fig. 3 and 4, the sublingual neostigmine bromide tablets prepared in example 1 and the sublingual neostigmine bromide films prepared in example 9 show substantially comparable dissolution rates and mucosal permeability.

The sublingual tablet or the sublingual membrane provided by the invention is applied to the preparation of the medicine for treating myasthenia gravis, postoperative abdominal distension and urine retention, supraventricular paroxysmal tachycardia treatment and detoxification when tubocurarine is excessive, and has huge clinical application prospects.

It should be noted that, as will be understood by those skilled in the art, in the present invention, the active ingredient is at least one of neostigmine, neostigmine bromide, neostigmine methosulfate, and pirstigmine bromide, and is not limited to neostigmine bromide in the above examples. In the present invention, mucosal formulations applied to other mucosal sites, such as buccal mucosa, sublingual mucosa, nasal mucosa, rectal mucosa, vaginal mucosa, can also be prepared, and are not limited to the sublingual mucosal route in the above examples.

In summary, the present invention provides a novel formulation of neostigmine and its derivatives. The preparation comprises sublingual mucosa preparation, nasal mucosa preparation, rectal mucosa preparation, vaginal and uterine mucosa preparation and buccal mucosa preparation which take neostigmine or neostigmine derivatives as active ingredients. The preparation is administrated through any mucous membrane route of buccal mucosa, sublingual mucosa, nasal mucosa, rectal mucosa or vagina and uterine mucosa, and has great clinical application value in preparing medicines for treating myasthenia gravis, postoperative abdominal distension and urinary retention, supraventricular paroxysmal tachycardia treatment and detoxification when tubocurarine is excessive.

Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the spirit and scope of the present invention.

Claims

1. A novel formulation of neostigmine and its derivatives characterized by: the novel preparation is a sublingual tablet or sublingual membrane, which takes neostigmine or neostigmine derivatives as active ingredients and also comprises a pH regulator and pharmaceutically acceptable auxiliary materials; in a single dose of the novel formulation, the active ingredient is present in an amount of 0.1mg to 20 mg.

2. Novel formulations of neostigmine and its derivatives according to claim 1, characterized in that: in the novel preparation, the preferable dosage of the active ingredient is 1 mg-15 mg; the dosage of the pH regulator is 5-15% (w/w).

3. Novel formulations of neostigmine and its derivatives according to claim 1, characterized in that: the acidity of the new preparation is 6-8; the pH regulator is sodium carbonate.

4. A novel formulation of neostigmine and its derivatives according to claim 2, characterized in that: the preferable dosage of the active ingredients is 1mg-5 mg; the preferable dosage of the pH regulator is 7-13% (w/w).

5. Novel formulations of neostigmine and its derivatives according to claim 4, characterized in that: the more preferable amount of the pH regulator is 9% to 11% (w/w).

6. Novel formulations of neostigmine and its derivatives according to claim 1, characterized in that: the sublingual tablet comprises 10-90 w/w% of a filling agent, 1-10 w/w% of a disintegrating agent, 20-60 w/w% of an acid-base effervescent agent and 0.5-2 w/w% of a lubricating agent, wherein the sum of the components is 100% (w/w).

7. Novel formulations of neostigmine and its derivatives according to claim 1, characterized in that: the sublingual film comprises 10.0-95.0 w/w% of water-soluble film forming materials, 5.0-30.0 w/w% of plasticizers, 0.05-1.5 w/w% of surfactants, and a proper amount of stabilizers, flavoring agents and opacifiers, wherein the sum of the components is 100% (w/w).

8. Novel formulations of neostigmine and its derivatives according to claim 1, characterized in that: the active ingredients include, but are not limited to, neostigmine bromide, neostigmine mesylate, pirstigmine bromide, or combinations thereof.

9. The application of neostigmine and a mucous membrane preparation of derivatives thereof is characterized in that: the mucosal preparation takes neostigmine or neostigmine derivatives as active ingredients, and the neostigmine or neostigmine derivatives can be applied to the preparation of medicines for treating myasthenia gravis, postoperative abdominal distension and urine retention, supraventricular paroxysmal tachycardia and detoxifying when tubocurarine is excessive through any one of the mucosal routes of buccal mucosa, sublingual mucosa, nasal mucosa, rectal mucosa or vagina and uterine mucosa;

the mucosal preparation comprises the novel preparation of neostigmine and the derivatives thereof as claimed in any one of claims 1 to 8, and nasal, rectal, vaginal and uterine mucosa and buccal mucosa preparations which take neostigmine or neostigmine derivatives as active ingredients.

10. A mucosal delivery method of neostigmine and its derivatives, characterized in that: the mucosal administration method comprises taking neostigmine or neostigmine derivatives as active ingredients, adopting the novel preparation of neostigmine and derivatives thereof as claimed in any one of claims 1 to 8 to carry out mucosal administration according to mucosal types, or adopting one of nasal mucosa preparation, rectal mucosa preparation, vaginal and uterine mucosa preparation and cavity buccal mucosa preparation which takes neostigmine or neostigmine derivatives as active ingredients as claimed in claim 9 to carry out mucosal administration.