Auxiliary material	% contents	Function
			Sodium caprate	40-80	Sorbefacient
Microcrystalline cellulose	15-75	Diluent
			Hydroxypropyl methyl cellulose	0-20	Matrix
Mannitol	0-30	Diluent
			Sodium carboxymethylcellulose	1-8	Disintegrant
PVP	1-5	Adhesive
			Superfine silica gel powder	1-2	Glidant

Magnesium stearate	0.1-2	Lubricant
			Medicine	0.1-20	Therapeutic component

In a particular embodiment, the amount of the especially sodium caprate of the sorbefacient in formula, can be 400mg, 300mg,200mg, 100mg, 50mg or 25mg.200mg, 100mg or 50mg are best, or even 100mg more preferable.

The presence of hydroxypropyl methyl cellulose (HPMC) can adjust sorbefacient and therapeutic component in label in labelRelease behavior.The HPMC of different viscosities can be added in label by different proportion, adjust the Adhesion property of label.

The usual potency of macromolecular drug of the present invention is higher, therefore the medicament contg in label is generally relatively low, such as1%, 0.1%, 0.01%.Significant challenge in terms of the quality that these low dosage formulas face includes stability, content uniformityAnd analysis method etc..It is generally necessary to design technology condition, the stability of holding therapeutic component while content uniformity is ensured.

Unlike low dose formula facing challenges, heavy dose of formulation of drug ratio is higher, and what is more faced isThe problems such as compatibility, mobility.

In instantiation, the medicine in formula can exist in the form of calcium phosphate nano particle, its specific descriptionReferring to U.S. Patent application 2005/0234114, Application U.S. Serial No 12/434,557 and PCT Application No. WO 2005/Described in 084637 and WO 2009/135190.Above-mentioned entirety is quoted.

The pharmaceutical agent combinations of the present invention can also be suppository form, pass through rectally.By component and nonirritating, normal temperatureIt is solid down, and is mixed under rectal temperature for the auxiliary material of liquid, after reaching rectum, medicament dissolves, insoluble drug release.These materialsIncluding but not limited to cocoa butter and polyethylene glycol etc..

The pharmaceutical agent combinations of the present invention can also be to be administered by vagina administration mode.Mode suitable for vagina administration include butIt is not limited to vagina and fastens agent, tablet or tampon agent.

Pharmaceutical agent combinations in invention can prepare each dosage unit and conveniently take, and can be prepared by known any method.

Bio-adhesive layer

As described above, bio-adhesive layer coating of the pharmaceutical agent combinations of the present invention using bio-adhesive polymer composition.OneIn a little instantiations, bio-adhesive coating directly contacts with solid pharmaceutical preparation.In other instantiation, bio-adhesive coatingThere is one or more layers intermediate layer between solid pharmaceutical preparation.It is non-to ooze or pellicle is located at solid pharmaceutical preparation and life in some instantiationsBetween thing adhesion layer, or, bio-adhesive layer can be located at solid pharmaceutical preparation and it is non-ooze or pellicle between.

In instantiation, bio-adhesive polymer can directly be attached to targeting moiety, such as intestines and stomach in bio-adhesive layerIn.Bio-adhesive polymer includes but is not limited to carbomer, polycarbophil, chitosan, sodium alginate, mercaptomerin sodium, gelatin, hydroxylPropyl methocel, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, polyvinyl alcohol, polyethylene glycol, poly- secondAlkene pyrrolidone, fumaric acid anhydride copolymer, polyester, acrylate, polysaccharide, modified dextrans, Sodium Hyaluronate, fruitGlue, xanthans, salt and its derivative and mixture.In instantiation, bio-adhesive layer includes hydroxypropyl methyl celluloseOr polycarbophil AA1 (HPMC).

In a particular embodiment, bio-adhesive layer is answered by what such as bio-adhesive polymer, plasticizer or other materials formedCondensation material forms, to adjust rate of release.Tend to the bio-adhesive material using high level.In a particular embodiment, it is rawThe ratio that thing sticks polymer is 50%, or higher than 65%, or higher than 75%, or higher than 80%, or higher than 90%.

In instantiation, bio-adhesive layer can contain enteric material, as cellulose acetate phthalate (S orL), HPMCP, hydroxypropyl methyl cellulose acetate, acetic acid -1,2,4 benzenetricarboxylic acid celluloses, polyvinyl acetate phthalate, methacrylic acid copolymer, shellac, salt, derivative and compound.

The presence of enteric material in bio-adhesive layer, can protect integrality of the preparation in gastric juice, and preparation reaches enteron aislePH increases, medicine start to discharge and absorbed afterwards.In order to keep stability of the preparation in acid, it is necessary to the enteric material of higher proportionMaterial, such as 50% even more high.Therefore, when enteric material as little as 10~20%, and it is very when can still keep the stability in acidIt is wonderful.

In a particular embodiment, the enteric material content in bio-adhesive layer 5~50%, 10~35%, best 15~25%.

In a particular embodiment, plasticizer can be contained in bio-adhesive layer, such as glycerine, glycerol acetate, polyvinyl alcohol, poly-Ethylene glycol, its derivative or mixture.In a particular embodiment, the plasticizer in bio-adhesive layer is 1~35%, 5~25%Preferably, 10~15% is more preferable.

In instantiation, percentage by weight of the bio-adhesive layer in pharmaceutical agent combinations can be 0.1%~10%, 1~5%, preferably 2~3%.

In a particular embodiment, the thickness of bio-adhesive layer can be adjusted to obtain preferable release dynamics parameter and suctionProduce effects fruit.

Enteric material can quickly dissolve in appropriate pH.Due to the enteric material in bio-adhesive layer and enteric layer in formulaThe content of material does not have marked difference, therefore the bio-adhesive layer containing enteric material should be consistent with the rate of dissolution of enteric layer's.

Therefore, with the recipe ratio containing enteric layer compared with when the formula for finding to contain identical enteric material in bio-adhesive layerAbsorption and pharmacology accelerate when, this is very unexpected.

This can increase enteric material addition bio-adhesive layer the absorption window in enteron aisle it is surprisingly found that showMouthful, clinical response is earlier, generally more desirable.

The formula of enteric material addition bio-adhesive layer is reduced into requirement and the technical process to coating, reduces productionTime, while reduce production cost.

Impermeable or semi-permeable layer

Ooze or semi-permeable layer can be used for adjusting release dynamics, further reduced to the need of sorbefacient as described above, non-Measure, improve the assimilation effect and bioavailability of the difficult component that absorbs the drug.It is non-to ooze or semi-permeable layer contains out in instantiationPassage is put, the therapeutic component and sorbefacient being easy in solid pharmaceutical preparation unidirectionally discharge.The size and shape of passage depends on solidThe property of body preparation and the release dynamics feature needed.The size of passage is generally can determine whether by veteran.In some toolsIn body example, passage has been fully contemplated by the side of tabletting or capsule.

In a particular embodiment, unidirectionally the release channel of release tabletting covers the 20~90% of the tabletting side gross area, or30~80%, best 50~70%.

The shape of open channel can be any shape, circle, rectangle, triangle, trapezoidal or square.

It is non-to ooze or semi-permeable layer contains one or more hydrophilic polymers or/and one or more hydrophobic in instantiationPolymer.Hydrophilic polymer includes but is not limited to, polymer (for example, gelatin and casein), pectin, agar based on albumen(agar), chitosan, carragheen, starch, glucan, methylcellulose, calcium carboxymethyl cellulose, sodium carboxymethylcellulose, friendshipJoin sodium carboxymethylcellulose polymer (such as AC-DI-SOL), microcrystalline cellulose, ethyl cellulose, hydroxy ethyl fiberElement, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalic acid, cellulose ether, acetate fiberElement, cellulose acetate phthalic acid, other cellulose derivatives, polyvinyl alcohol, polyvinylpyrrolidone (PVP), crosslinking are poly-Tie up ketone, other polyvinyls and copolymer, guar gum, poloxamer, polyethylene glycol, polyethylene glycol oxide, polyacrylic acid,Polyethers, alkoxy polymers, sodium alginate, xanthans, other natural hydrogels obtain gel from natural prodcuts, or bothMixture.The example of hydrophobic polymer includes but is not limited to, and (such as polycaprolactone (PCL), polyamides are sub- for ethyl cellulose, polyesterAmine (PEA), polyhydroxyalkanoate (PHA), PLA (PLA), PLA alcohol copolymer (PLGA), poly butyric ester-Hydroxyl valerate (PHBV) and polybutadiene acid esters (PBSA), wax, low melt wax, polyethylene and ethylene copolymer, ethene/acetic acid secondAlkene, polypropylene, polyurethane, ethylene/vinyl alcohol, polyvinyl alcohol, Corvic, polyolefin or any combinations.

It is non-to ooze or semi-permeable layer also includes solvent or plasticizer in instantiation, so that coatings keep flexible.Solvent bagInclude it is any oozed or solvent that semipermeable membrane material is compatible with non-, including such as water and ethanol.Plasticizer includes but unlimited and citric acidPolyalkylene oxides (such as polyethylene glycol, poly- the third two of ester (for example, triethyl citrate, triacetyl glycerine), low molecule amountAlcohol, polyethylene/propyleneglycoles), glycerine, pentaerythrite, monoglyceride, oxalic acid or triacetate, propane diols, butanedioic acid twoEthyl sodium, sugar alcohol and corn syrup and composition.

In instantiation, non-infiltration layer is by ethyl cellulose and glycerine, polyethylene glycol or acetoglyceride as plasticizerComposition.In some instantiations, semi-permeable layer is by cellulose acetate and polyethylene glycol, glycerol acetate or glycerine as plasticizerComposition.

In a particular embodiment, it is non-ooze or pellicle in plasticizer content 5~40%, or 10~30%, 15~25% is best.

It is non-to ooze or the weight of semi-permeable layer accounts for 0.1~10% of gross weight or so, or 1~5% or so in instantiation, mostFortunately 2~3% or so.

Can further add other compositions in non-infiltration layer, as preservative, protective agent, other can improve stability of drug products intoGrade.Addible plasticizer and component visible M.＆I.Ash, T_HE H_{ANDBOOK OF} P_{HARMACEUTICAL} A_DDITIVES (3^rdEd.,Synapse Information Resources, Inc., 2007).

In a particular embodiment, unidirectional release channel can be prepared by laser ablation process, and the technique has been used for permeatingPump produces.But unlike osmotic pumps tabletting, bigger (the unidirectional release channel a diameter of 3 of 10mm tablettings of unidirectional release channel~9mm, and osmotic pumps are only 0.5mm), coatings are thinner, therefore, it is necessary to using the different lasing light emitters and dress for adapting to the present inventionPut.

Enteric layer

In instantiation, the pharmaceutical agent combinations containing label and the coating of bio-adhesive layer and unidirectional releasing layer in the present invention are alsoFurther using site selection material coating, allow medicament to discharge in the position that intestines and stomach are specified.Site selects material by pHSensitive material forms, and is dissolved within the border in certain pH range ring.Using site selection material coating, sticking for carrier selectivity can be madeAttached layer is exposed to the specified location of intestines and stomach.

Enteric coating polymer may be selected, but be not limited to cellulose acetate-phthalate (S or L), hydroxylPropyl methocel phthalic acid ester, hydroxypropyl methyl cellulose amber acetic acid, acetic acid-TMLA cellulose,Polyvinyl acetate phthalate, methacrylic acid copolymer, shellac, its salt, derivative and compound etc..In instantiationIn, enteric-coating material refers toL30D-55。

In instantiation, site selection material includes selectivity and is attached to colon or the material in colon release.It is thisMaterial includes but is not limited to azobenzene polymer, colon degradation of polysaccharide such as pectin, starch, guar gum, xylan, cyclodextrin, PortugalGlycan, its salt and derivative etc..

The rate of release that the thickness of coating is as needed selects, and this relies on the property and thickness with coating.In instantiationIn, enteric layer weight accounts for about the 1~15% of solid pharmaceutical preparation gross weight, 3~12%, best 6~10%.

Preparation method

The preparation method of medicament composition provided by the invention comprises the following steps：Prepare the treatment group for including effective dosePoint, the solid pharmaceutical preparation of sorbefacient and pharmaceutical excipients；Using bio-adhesive polymer coating bio-adhesive layer, optional non-oozeOr semi-permeable layer coating, there is open channel, therapeutic component and sorbefacient is unidirectionally discharged from preparation.In instantiationIn, bio-adhesive layer and it is non-ooze or the coating of semi-permeable layer order be reversible.In instantiation, preparation method includes furtherIt is coated using enteric layer.

In instantiation, can use bio-adhesive material and it is non-ooze or semi-permeable material such as ethyl cellulose orL30D-55 water slurry.In some instantiations, the material can directly be allocated without using solvent to be madeWith.In some instantiations, use can be directly adjusted after polycaprolactone (PCL) or wax heating.

In a particular embodiment, pharmaceutical agent combinations of the invention are more likely to be prepared as the form of tabletting or capsule.Tabletting pieceCore generally use interlocks tabletting and prepared.Tabletting preparation technology, including wet granulation and direct tablet compressing, have been widely used(Developing Solid Oral Dosage Forms：Pharmaceutical Theory and Practice.Ed.byQiu et al.Academic Press 2009)。

However, the compression forms of macromolecular drug are still there is challenge.Most of macromolecular drugs are polypeptide and albumen,Complicated, stability is very poor.The usual potency of these macromolecular drugs is very high, therefore dosage is relatively low, and this is just needed in technique workSuch as homogeneity (the Formulation and Analytical Development of holding content in wet-granulation process in journeyfor Low Dose Oral Drug Products.Ed.by Zheng.Wiley 2009)。

In process exploitation engineering, Exenatide and insulin all suffer from this problem.When the wet granulation technology system of useDuring standby tabletting, it is found that the homogeneity of content meets the requirements, but Exenatide and unstable, and Ai Saina under identical conditionPeptide material powder can keep stable after directly being mixed with auxiliary material in formula.

Therefore, direct tablet compressing is another the optional technique for preparing the medicine that acceptable dosage is 0.5~2mg.GenerallyThe direct tablet compressing technique of low-dose drugs is relatively easy, because the content of medicine is very low, the performance for not interfering with tabletting is for example compatibleProperty, the hardness of mobility and tabletting, as long as ensureing the homogeneity of content.Medicine such as insulin powder etc., for the small of crystallizationParticle, hygroscopicity is poor, as long as the particle diameter of suitable control drug powder, you can tabletting is prepared using direct tablet compressing technique,

Recipe ingredient in the present invention, due to introducing the sorbefacient such as sodium caprate of larger proportion, its compatibility compared withDifference, while particle agglomeration may be caused, therefore there is other challenge.In a particular embodiment, the direct pressure in the present inventionSlice prescription is as shown in the following chart.

Auxiliary material	% contents	Function
			Sodium caprate	40-80	Sorbefacient
Microcrystalline cellulose	15-75	Diluent
			Hydroxypropyl methyl cellulose	0-10	Matrix
Sodium carboxymethylcellulose	1-8	Disintegrant

Superfine silica gel powder	1-2	Glidant
			Magnesium stearate	0.1-2	Lubricant
Medicine	0.1-20	Therapeutic component

The medicine of Exenatide class, it is necessary to unit dose it is even lower, such as/or drug powder there is hygroscopicity, due toIt is difficult to reach content uniformity in preparation process, therefore is not proposed with direct tablet compressing technique (ibid. of Zheng 2009).

In the present invention, by using non-solvent granulating process, it can keep between medicine stability and content uniformityBalance.In a particular embodiment, drug powder is suspended in the undissolved fluid media (medium) of medicine, by ultrasound or is homogenized and can be controlledMake and adjust the particle diameter of drug particles.Adhesive can be added in suspension to increase uniformity.What suspension was generally commonly used togetherPelletization is similar, adds the mixture of each auxiliary material.

Non-solvent granulating process in the present invention, the stability of medicine can be kept while content uniformity is ensured.The PEG400 of low molecule amount reduces the stability of Exenatide, and content uniformity is fine.

In a particular embodiment, non-solvent usually used in the present invention includes ethanol, isopropanol, acetone and acetic acid secondEster.

Application method

The pharmaceutical agent combinations of the present invention can be effectively by drug delivery to the mucomembranous surface specified.When pharmaceutical agent combinations selectivityWhen being attached to mucomembranous surface, therapeutic component and sorbefacient in solid pharmaceutical preparation can unidirectionally be discharged into mucomembranous surface simultaneously, togetherWhen produce high local concentrations.So, in drug regimen of the invention, the sorbefacient of relatively low amount will be obviously promoted medicineAbsorb, be otherwise invalid.

Therefore, on the one hand, the invention provides a kind of method that medicine is transmitted using pharmaceutical agent combinations.On the other hand, this hairThe bright patient to need provides the method for drug therapy, including takes pharmaceutical agent combinations disclosed by the invention to patient part, especiallyIt is mucomembranous surface.Pharmaceutical agent combinations can be administered by known any mode, include but is not limited to oral, oral cavity, sublingual administration,Vagina, rectally.Can whole body or local use.

Brief description of the drawings

Fig. 1 show using hydroxypropyl methyl cellulose (HPMC) andAfter L30D-55 enteric coatings, in tablettingExenatide and sorbefacient sodium caprate be synchronous release.

Fig. 2A and 2B shows using HPMC and enteric coating and cellulose acetate is respectively adopted and ethyl cellulose is unidirectionally releasedPut the influence discharged after coating to Exenatide in tabletting and sodium caprate.

Fig. 3 A and 3B show bio-adhesive HPMC, polycarbophil AA1 or chitosan coat to Exenatide bioavailabilityInfluence, sodium caprate addition is respectively 100mg and 400mg in tabletting, enteric coating.

After Fig. 4 shows AA1 and enteric coating, influence of the sodium caprate addition to Exenatide bioavailability.

Fig. 5 shows that under the conditions of 50mg sodium caprates, HPMC and enteric coating cellulose acetate and ethyl cellulose are unidirectionally releasedPut influence of the coatings to Exenatide bioavailability.

Fig. 6 shows under the conditions of 100mg sodium caprates, HPMC, ethyl cellulose one-way membrane and enteric coating, oral Ai SainaThe relatively hypodermic bioavailability of peptide.

Fig. 7 shows that under 200mg sodium caprates, HPMC, cellulose acetate one-way membrane coating conditions enteric layer is to ExenatideThe influence of bioavailability.

Fig. 8 A show the insulin tabletting containing 200mg sodium caprates, HPMC coatings and unidirectional releasing layer coating in 0.01NRelease dynamics in HCl (2h) and pH6.8 simulated intestinal fluids (3~7h)；Fig. 8 B and Fig. 8 C show oral insulin to life respectivelyThe influence of the blood glucose and serum insulin of long chalone infusion dog.

Fig. 9 shows that two kinds of oral insulin formulas are transfused the hypoglycemic effect of dog to growth hormone release inhibiting hormone.The first formula is two layersTabletting, contain 200mg sodium caprates, HPMC coatings and unidirectional releasing layer coating.Second of formula is three layers of coating, contains 200mgSodium caprate, HPMC coatings, the coating of unidirectional releasing layer and enteric layer coating.

Figure 10 shows stability of the Exenatide in solid pharmaceutical preparation, different formulations be individually positioned in 60 DEG C (Figure 10 A),0,5,10 day under the conditions of 92.5% humidity (Figure 10 B), 4500lux illumination (Figure 10 C).

Figure 11 shows influence of the oral insulin to normal dogs blood glucose.Experiment formula used is three layers of coating tabletting, is contained100mg sodium caprates, HPMC coatings, unidirectional releasing layer and enteric coat layer.

Figure 12 shows what the unidirectional release channel that manual pasting technique and laser ablation process are formed absorbed to ExenatideInfluence.It is two-layered coating tabletting used in experiment, the HPMC containing 200mg sodium caprates, containing L30D-55 is coated, using manual or sharpThe unidirectional release channel that light technique is formed.

Figure 13 shows that oral insulin dosage (25U, 25U × 2,50U) is transfused the hypoglycemic effect of dog to growth hormone release inhibiting hormone.ExperimentUsed is two-layered coating tabletting, HPMC coatings and unidirectional releasing layer containing 200mg sodium caprates, containing L30D-55.

Figure 14 shows that bio-adhesive polymer content is in Exenatide release dynamics and normal dogs in bio-adhesive layerThe influence of absorption.It is two-layered coating tabletting used in experiment, containing 100mg sodium caprates, containing L30D-55's and 65% or 80%HPMCThe unidirectional releasing layer that coatings and laser ablation are formed.

Embodiment

The present invention is expanded on further below by the detailed description of the specific embodiment to the present invention, but embodiment is notLimitation of the present invention.Declare at the same time：On the basis of without departing substantially from the application original meaning and scope, industry professional person also may be usedTo derive various system under the inspiration of the application, the application covers all equivalent or similar systems.

Embodiment 1

The preparation of tabletting

The preparation of 1.1 tabletting labels

Tabletting label is prepared by the proportioning of table 1, is suppressed after each component is well mixed using monolithic tablet press machine.Except Exenatide andOutside magnesium stearate, remaining component is weighed and is well mixed in advance.Made again with 15% PVP ethanol solutions (25%) for adhesiveGrain, 60 DEG C of dry 2hr.Particle crosses 22 mesh sieves and weighs particle, Exenatide and magnesium stearate according to monolithic aequum.Each componentIt is tabletted after well mixed.Weigh, the label more than average weight ± 5% is cast out.

Table 1：Exenatide and sodium caprate proportioning (unit in label：mg)

1.2 bio-adhesive layers

Label is further coated using bio-adhesive polymer such as hydroxypropyl methyl cellulose or polycarbophil AA1.UsingWhen HPMC is coated, HPMC is formulated as 2% aqueous solution, and (BY300A, Huanghai Sea instrument) is coated in small-sized seed-coating machine afterwards,Coating weight gain amount is 2% or so of label.During AA1 coatings non-using card ripple, the non-AA1 of card ripple is formulated as 4% ethanol solution, togetherSample operates in small-sized seed-coating machine, gain in weight 3%.Tabletting after coating is dried 14 hours at 30 DEG C.

1.3 unidirectional releasing layers

Label is then coated using non-or semi-permeable material.During operation, first by the use of sticky paper using the side of label cover asThe passage that medicine unidirectionally discharges, then using 4% ethyl cellulose ethanol solution (wherein containing 20% glycerol acetate as increasingMould agent) or 3% cellulose acetate (using acetone and formic acid as solvent, 9: 1V/V, containing 20% polyethylene glycol 2000 as be plasticizedAgent) solution coating.After coating, the dry 30min of 30 DEG C of tabletting, the gain in weight of coating is adjusted to 2~5%.Sticky paper is peeled off afterwards,Expose release channel.

1.4 enteric layer

Tabletting uses enteric polymerL30D-55 is coated.Coating solution includes 200g L30D-55,12g talcumsPowder, 6g Macrogol 6000s, adjusted after water mesoscale eddies is uniform to 400ml, it is standby.Coating is carried out in small-sized seed-coating machine, is wrappedClothing gain in weight is the 10% of tabletting gross weight.Tabletting is dried 14 hours in 30 DEG C.

Embodiment 2

The synchronous release of Exenatide and sodium caprate

The in-vitro evaluation release behavior of Exenatide and sodium caprate in different formulations.

Stability of the preparation in acid is primarily looked at.The tabletting of enteric coating is put into 100ml 0.1N hydrochloric acid,37 DEG C vibrate 2 hours in dissolution rate instrument, observe the stripping property of medicine.Different time samples, in HPLC (Waters) test solutionExenatide and capric acid na concn.As a result show that tabletting is kept completely in acid, no Exenatide and capric acid sodium leakage.

Tabletting is moved into 100ml simulated intestinal fluids (pH 6.8) afterwards, 37 DEG C of vibrations, different time sampling, investigates tablettingThe release behavior of middle Exenatide and sodium caprate.Table 2 lists sodium caprate containing 50mg and is coated HPMC layers and enteric layer successivelyThe release data of Exenatide and sodium caprate in tabletting.Release profiles are shown in Fig. 1.It can be seen that under the conditions of pH6.8, Exenatide andThe release of sodium caprate is almost synchronous.

The Exenatide of table 2 and sodium caprate are from the release in the tabletting of HPMC and enteric coating

Time (minute)	Exendin discharges (%)	Certain herbaceous plants with big flowers acid sodium discharges (%)
			0	0	0
15	0	0
			30	0	0
60	21.73	23.54
			90	66.91	60.48
120	90.15	91.55
			180	92.82	92.36
240	92.85	87.72

Table 3 lists sodium caprate containing 50mg and is coated successively in the tabletting of HPMC layers, layer of cellulose acetate and enteric layer and endsFill in the release data of that peptide and sodium caprate.Release profiles are shown in Fig. 2A.It can be seen that the release of Exenatide and sodium caprate is also almost sameStep.But compared with the tabletting of uncoated layer of cellulose acetate, release time is obviously prolonged.

The Exenatide of table 3 and sodium caprate from coating HPMC, enteric layer, layer of cellulose acetate tabletting in release

Time (minute)	Exenatide discharges (%)	Sodium caprate discharges (%)
			0	0	0
15	0	0
			30	0	0
60	6.05	7.3
			90	11.41	19.75
120	27.91	44.4
			180	66.99	85.07
240	74.01	91.34

Table 4 list sodium caprate containing 50mg and be coated successively HPMC layers, ethylcellulose, enteric layer tabletting releaseData.Release profiles are shown in Fig. 2 B.It can be seen that as with previous experiment, the release of Exenatide and sodium caprate is also almost synchronization.It is worth noting that, using ethylcellulose coat tabletting release than using cellulose acetate coating tabletting release whenBetween more extend.

The Exenatide of table 4 and sodium caprate are from the release in the tabletting for being coated HPMC, enteric layer, ethylcellulose

Time (minute)	Exenatide discharges (%)	Sodium caprate discharges (%)
			0	0	0
15	0	0
			30	0	0
60	0	0
			90	11.43	7.96
120	25.65	20.78
			180	37.63	40.05
240	49.75	51.29
			300	76.22	87.28
360	79.85	87.58
			540	79.64	91.7

Embodiment 3

Bio-adhesive layer to Exenatide beasle dog body absorption influence

Using the influence that is absorbed to Exenatide of healthy beasle dog evaluation different formulations, capric acid containing 100mg respectively in formulaSodium is as sorbefacient.

Healthy beasle dog 12,8~12kg of body weight is chosen, nursing is responsible for by Shanghai Univ. of Traditional Chinese Medicine's animal center.Will compareLattice dog is randomly divided into different treatment groups, and animal has the mastery phase of one week or so after experiment.During experiment, beasle dog overnight fasting,10ml water will be gavaged after the direct feeding of tabletting.Feeding after testing 6 hours.

Beasle dog is tested and packet situation is shown in Table 5.

Animal packet situation in experiment of the bio-adhesive layer of table 5 to Exenatide inhalation effects

Numbering	Group	Administrations
			1	Blank group	Comfort piece
2	Hypodermic injection group	60 μ g/10mM sodium acetate solutions, pH4.0 is subcutaneously injected
			3	Without adhesion layer group	3mg Exenatides, 100mg sodium caprates, enteric coating
4	HPMC coating groups	3mg Exenatides, 100mg sodium caprates, HPMC and enteric coating
			5	AA1 coating groups	3mg Exenatides, 100mg sodium caprates, AA1 and enteric coating
6	Chitosan Coating group	3mg Exenatides, 100mg sodium caprates, chitosan and enteric coating

After administration, different time points using test tube of hepari take blood vessel take blood 1.5ml, 3000rpm take 0.5 after centrifuging 10 minutes~0.6ml serum.- 20 DEG C of blood serum sample freezes, and Exenatide concentration is using enzyme linked immunological kit test (PhoenixPharmaceuticals, Inc.).

Sample includes testing after standard items dilute 5 times.Exenatide concentration in blood serum sample is calculated according to standard curve,According to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Table 6 and Fig. 3 A.

During 6 sodium caprate containing 100mg of table, influence of the bio-adhesive layer to Exenatide bioavailability

Group	Number of animals	Bioavailability (%)
			Blank	3	0
It is subcutaneously injected	3	100
			Without adhesion layer	3	0.19
HPMC is coated	3	2.12
			AA1 is coated	3	2.37
Chitosan Coating	3	1.25

As a result show, bio-adhesive layer can significantly affect the absorption of Exenatide.During sodium caprate containing 100mg, such as inanimate objectAdhesion layer, the absorption of Exenatide is very low, and Ai Saina is significantly improved after adding the non-AA1 of card ripple, chitosan or HPMC coatingsThe absorption of peptide.

Embodiment 4

The influence (sodium caprate containing 400mg) that bio-adhesive layer absorbs to Exenatide in beasle dog body

When evaluating sodium caprate containing 400mg using healthy beasle dog, influence that different formulations absorb to Exenatide.

Beasle dog is tested and packet situation is shown in Table 7.

The bio-adhesive layer of table 7 is to the animal packet situation in the experiment of Exenatide inhalation effects

Numbering	Group	Administrations
			1	Blank group	Comfort piece
2	Hypodermic injection group	60 μ g/10mM sodium acetate solutions, pH 4.0 is subcutaneously injected
			3	Without adhesion layer group	5mg Exenatides, 500mg sodium caprates, enteric coating
4	AA1 coating groups	3mg Exenatides, 400mg sodium caprates, AA1 and enteric coating

Sample includes testing after standard items dilute 5 times.Exenatide concentration in blood serum sample is calculated according to standard curve,According to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Table 8 and Fig. 3 B.

During 8 sodium caprate containing 400mg of table, influence of the bio-adhesive layer to Exenatide bioavailability

Group	Number of animals	Bioavailability (%)
			Blank group	3	0
It is subcutaneously injected	3	100
			Without adhesion layer	3	0.61
AA1 coating groups	3	2.34

Embodiment 5

The influence that sodium caprate addition absorbs to Exenatide in beasle dog body

The influence absorbed using being added in healthy beasle dog evaluation formula after not same amount sodium caprate to Exenatide.

Beasle dog is tested and packet situation is shown in Table 9.

Animal packet situation in the experiment of the sodium caprate addition Exenatide inhalation effects of table 9

Numbering	Group	Administrations
			1	Blank group	Comfort piece
2	Hypodermic injection group	60 μ g/10mM sodium acetate solutions, pH 4.0 is subcutaneously injected
			3	50mg groups	3mg Exenatides, 50mg sodium caprates, AA1 and enteric coating
4	100mg groups	3mg Exenatides, 100mg sodium caprates, AA1 and enteric coating
			5	200mg groups	3mg Exenatides, 200mg sodium caprates, AA1 and enteric coating
6	400mg groups	3mg Exenatides, 400mg sodium caprates, AA1 and enteric coating

Sample includes testing after standard items dilute 5 times.Exenatide concentration in blood serum sample is calculated according to standard curve,According to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Table 10 and Fig. 4.

Table 10 contains in HPMC and enteric coating tabletting, influence of the sodium caprate addition to Exenatide bioavailability

Group	Number of animals	Bioavailability (%)
			Blank group	3	0
Hypodermic injection group	3	100
			50mg groups	3	0.08
100mg groups	3	2.37
			200mg groups	3	2.62
400mg groups	3	2.34

As a result show that the absorption of Exenatide is relevant with the addition of sodium caprate, add 50mg sodium caprates as absorption enhancementDuring agent, the absorption of Exenatide is very weak.When the addition of sodium caprate is higher than 100mg, the absorption of Exenatide significantly improves, and entersOne step increases the addition of sodium caprate, and the absorption of Exenatide does not significantly improve.

Embodiment 6

Unidirectional delivery formulations to Exenatide beasle dog body absorption influence

Using the absorption of Exenatide after addition 50mg sodium caprates in healthy beasle dog evaluation formula.

Beasle dog is tested and packet situation is shown in Table 11.

11 unidirectional releasing layer of table influences the animal packet situation in experiment on bioavailability

Sample includes testing after standard items dilute 5 times.Exenatide concentration in blood serum sample is calculated according to standard curve,According to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Table 12 and Fig. 5.

The sodium caprate containing 50mg of table 12, in the tabletting using HPMC coatings, the unidirectional film that discharges is to Exenatide bioavailabilityInfluence

Group	Number of animals	Bioavailability (%)
			Blank group	3	0
Hypodermic injection group	3	100
			Without unidirectional coating group	3	0.08
CA coating groups	3	0.34
			EC coating groups	3	0.78

As a result show that the absorption of Exenatide can be carried further using after unidirectional release coating and bio-adhesive coatingIt is high.Coatings are not discharged unidirectionally, and the drug absorption and bioavailability of sodium caprate containing 50mg and HPMC coating tablettings are very low.Semi-permeable layer coating is carried out using cellulose acetate or non-infiltration layer coating is carried out using ethyl cellulose, and reserves and releases in wafer surfaceAfter putting passage, under same sodium caprate addition and HPMC coating conditions, the absorption of Exenatide is remarkably reinforced.

Embodiment 7

Using the absorption of Exenatide after addition 100mg sodium caprates in healthy beasle dog evaluation formula.

Beasle dog is tested and packet situation is shown in Table 13.

During 13 sodium caprate containing 100mg of table, unidirectional releasing layer influences the animal packet situation in experiment on bioavailability

Sample includes testing after standard items dilute 5 times.Exenatide concentration in blood serum sample is calculated according to standard curve,According to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Table 14 and Fig. 6.

The sodium caprate containing 100mg of table 14, using the biology profit of HPMC, unidirectional release film and enteric coating Exenatide tablettingWith rate

Group	Number of animals	Bioavailability (%)
			Blank group	3	0
Hypodermic injection group	3	100
			EC coating groups	3	4.98

As a result show the Exenatide tablet of the sodium caprate containing 100mg using HPMC coatings, ethylcellulose coat and intestinesAfter molten coating, the bioavilability of Exenatide is 4.98%.Tablet is coated using the ethyl cellulose of impermeability, andAfter release channel is reserved in side, significantly improve absorption and the bioavailability of Exenatide, and use same amount sodium caprate andThe bioavailability of the tablet of HMPC art for coating is only 2.37% (see embodiment 5).

Embodiment 8

Enteric layer to Exenatide beasle dog body absorption influence

It is prepared by 8.1 labels

Label is matched by table 15, is suppressed using monolithic tablet press machine.In addition to Exenatide and magnesium stearate, remaining component is advanceWeigh and be well mixed.It is granulated, is dried in vacuum overnight for adhesive with 15% PVP ethanol solutions (25%) again.Particle crosses 22Mesh sieve simultaneously weighs particle, Exenatide and magnesium stearate according to monolithic aequum.It is tabletted after each component is well mixed.MonolithicWeigh, the label more than average weight ± 5% is cast out.

Table 15：Each group distribution ratio (unit in label：mg)

8.2 bio-adhesive layers

Label is coated using HPMC, can be with or without enteric material in HPMCL30D-55.For withoutL30D-55 HPMC coating, using the aqueous solution containing 6%HPMC and 1.5%PEG6000 in seed-coating machine (BY300A, Huanghai Sea instrumentDevice) coating, the gain in weight of bio-adhesive layer is 3~4mg.Be coated for the HPMC containing L30D-55, using containing 3%HPMC and0.6%L30D-55 aqueous solution coating.40 DEG C of dryings 14 hours after compress tablet coating, adhesion layer weightening is 3mg.

8.3 unidirectional releasing layers

Further it is coated after the coating of tabletting bio-adhesive layer using semi-permeable cellulose acetate.During operation, use firstSticky paper covers the side of label in the passage as medicine, then using 3% cellulose acetate and 1.2% polyethylene glycol 2000(using acetone and formic acid as solvent, 9: 1V/V) solution is coated.40 DEG C of tabletting is dried overnight, and the gain in weight of coating is 2mg.Finally willSticky paper is peeled off, and exposes release channel.

8.4 enteric layer

Tabletting without L30D-55 coatings in adhesion layer is further coated using enteric material.Coating solution includes200gL30D-55,12g talcum powder, 6g Macrogol 6000s, adjusted after water mesoscale eddies is uniform to 400ml.Coating is in small packageCarry out in clothing machine, dried 14 hours in 40 DEG C of tabletting.Coating weight gain amount is 25mg.

Absorption of 8.5 Exenatides in beasle dog body

Using the influence that is absorbed to Exenatide of healthy beasle dog evaluation enteric layer, sodium caprate containing 200mg respectively in formula.

Healthy beasle dog 12,8~12kg of body weight is chosen, nursing is responsible for by Shanghai Univ. of Traditional Chinese Medicine's animal center.Will compareLattice dog is randomly divided into different treatment groups, and animal has the mastery phase of one week or so after experiment.Before experiment, beasle dog overnight fasting,Feed is limited during experiment.

Beasle dog is tested and packet situation is shown in Table 16.

During 16 sodium caprate containing 200mg of table, enteric layer influences the animal packet situation in experiment on bioavailability

Sample is diluted to containing test concentrations after 20% serum including standard items.Calculated according to standard curve in blood serum sampleExenatide concentration, according to the relative bioavailability of TG-AUC com-parison and analysis sample, data are shown in Fig. 7.

As a result show, the Exenatide tabletting containing 200mg sodium caprates, be respectively adopted HPMC, cellulose acetate coating orAfter HPMC, cellulose acetate and enteric coating, there is identical assimilation effect.Compared with the tablet of enteric layer coating, do not haveThe tablet maximum absorption band time of occurrence for having enteric layer to be coated shifts to an earlier date 1 hour.But either under absorption maximum concentration or curveArea, all do not significantly affected by enteric layer.Therefore, although employing enteric layer coating in some instantiations,It is not the key component of the present invention.

The drug absorption and clinical response occurred in two-layered coating formula shifts to an earlier date, and is preferably in clinical setting.

Embodiment 9

Oral insulin is transfused the effect of dog to growth hormone release inhibiting hormone

9.1 carry the preparation of insulin calcium phosphate nanoparticles

Using calcium phosphate nanoparticles as insulin carrier.Insulin is dissolved in 40ml solution As (5mg/ml, phosphorus containing 20mMAcid dihydride sodium, the HEPES buffer solutions of 20mM pH 6.9,2%PEG6000,0.5% urso UDCA, UDCA are molten before addingIn 1.5ml ethanol).The solution B (40ml 0.01N HCl CaCl2 containing 60mM) of same volume forms particle after being mixed with A.GrainSub- 15000rpm centrifuges 30min, vacuum drying.

Particle is suspended in pH 9.1 50mM phosphate buffers (0.2mg/ml), used after 37 DEG C of vibration 15minRP-HPLC surveys insulin content in particle.Content is calculated according to standard items.Insulin content is 0.56mg/mg in this batch, pancreasThe island element rate of recovery is 99%.

It is prepared by 9.2 labels

Matched according to table 16, using monolithic tablet press machine compressed cores.In addition to insulin, superfine silica gel powder and magnesium stearate, itsRemaining component is weighed and is well mixed in advance.Use 8% PVP ethanol solutions (25%) to be granulated for adhesive, be dried in vacuum overnight.Particle crosses 22 mesh sieves and adds insulin, superfine silica gel powder and magnesium stearate.It is tabletted after each component is well mixed.More than averageThe label of weight ± 5% is cast out.The hardness of label, thickness, friability are detected simultaneously.

Table 16：Piece core component (unit：mg)

9.3 bio-adhesive layers

Label uses HPMC E50 and enteric materialL30D-55 is coated.Prepare 3%HPMC and 0.6%L30D-55 suspension (BY300A, Huanghai Sea instrument) in seed-coating machine is coated.40 DEG C of dryings 14 hours after coating, adhesion layer weightening are3mg。

9.4 unidirectional releasing layers

Tabletting is further coated using semi-permeable cellulose acetate.First with the sticky papers of circular 7mm diameters by 9mmThe side of the tabletting label of diameter covers, then use 3% cellulose acetate and 1.2% polyethylene glycol 2000 (with acetone andFormic acid is solvent, 9: 1V/V) solution coating.40 DEG C of tabletting is dried overnight, and the gain in weight of coating is 2mg.Finally sticky paper is peeled off,As unidirectional release channel.

9.5 insulin releasing

Tabletting is put into dissolution rate instrument first, adds 100ml 0.01N HCl, keeps 37 DEG C and 100rpm release 2h,RP-HPLC is sampled in 1h and 2h respectively and surveys concentration.Tabletting is taken out to the simulated intestinal fluid SIF for being put into 100ml pH 6.8 afterwardsIn, discharged with the conditions of, different time sampling RP-HPLC surveys concentration.

Insulin releasing curve is shown in Fig. 8 A.Tabletting is stable in acid, and burst size is less than 10% in 2h.Release in SIFGently, 3~4h releases finish.

Effect of 9.6 tablettings to the dog of infusion growth hormone release inhibiting hormone.

The assimilation effect of oral insulin is evaluated using the beasle dog of venoclysis growth hormone release inhibiting hormone (1 μ g/kg/min).InfusionGrowth hormone release inhibiting hormone can suppress endogenic glucagons and insulin secretion (Sakurai et al.J.Clin.Investigation 54：1395,1974).Quick increase after blood sugar level initial reduction.This model can evaluate oral insulin simultaneouslyDrug effect and medicine generation.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.Growth hormone release inhibiting hormone (1 μ g/kg/min) is transfused by the ductus venosus of sacculus infusion pump, after being transfused 4h10ml water feeding tablettings.Feed is limited during experiment, different time points test tube of hepari takes blood vessel to take blood.Beasle dog is tested and packetIt is shown in Table 17.

Table 17：The oral insulin experimental animal packet of growth hormone release inhibiting hormone infusion dog

Group	n	Administration
			Blank group	3	Comfort piece
Hypodermic injection group	3	3.5U/ dogs, it is dissolved in 0.01N HCl
			Oral tabletting group	3	50U is oral

Using blood glucose meter and corresponding test paper (Johnson＆Johnson, OneTouch) test blood glucose.Blood sample 3000rpm fromThe heart 10 minutes, takes 0.5~0.6ml serum.- 20 DEG C of blood serum sample freezes, using ELISA kit (Linco) test sera pancreasIsland element concentration.Fig. 8 B and 8C are shown in influence of the oral insulin to blood glucose and serum insulin concentration respectively.

The animal blood glucose initial reduction of blank tabletting group, increases afterwards.Injection group blood glucose significantly quickly reduces.Oral pancreas isletThe blood glucose of plain tabletting group is also to significantly reduce.By calculating the AUC of blood glucose, the bioavilability of sample is 5%.

In experimentation, the serum insulin level of blank group is suppressed, and the serum insulin concentration of injection group is thenIt is significantly increased.The serum islet of oral insulin tabletting group is known as certain increase, and bioavilability is 2% or so.Confirm oralInsulin is similar to the insulin of hepatic portal or natural secretion, has higher bioavilability, and the insulin level of periphery is thenIt is relatively low, caused hypoglycemia this pressing issues in clinical practice can be significantly reduced.

Embodiment 10

The effect for the dog that the oral insulin tabletting of two-layered coating is transfused to growth hormone release inhibiting hormone

10.1 prepared by label

Matched according to table 18, using monolithic tablet press machine compressed cores.In addition to insulin, superfine silica gel powder and magnesium stearate, itsRemaining component is weighed and is well mixed in advance.Use 8% PVP ethanol solutions (25%) to be granulated for adhesive, be dried in vacuum overnight.Particle crosses 22 mesh sieves and adds insulin, superfine silica gel powder and magnesium stearate.It is tabletted after each component is well mixed.Detect labelHardness, thickness, friability.

Table 18：Piece core component (unit：mg)

10.2 bio-adhesive layer

Label is using HPMC E50 or adds addition enteric material simultaneouslyL30D-55 is coated.For not havingL30D-55 bio-adhesive layer coating, prepare 6%HPMC and 1.5%PEG6000 solution, coating pan (BY300A, Huanghai Sea instrumentDevice) in be coated.3~4mg of adhesion layer weightening.Coating for adding L30D-55, prepares 3%HPMC's and 0.6%L30D-55Suspension is coated in seed-coating machine.40 DEG C of dryings 14 hours after coating, adhesion layer weightening is 3mg.

10.3 unidirectional releasing layer

Tabletting is further coated using semi-permeable cellulose acetate.First will using the sticky paper of circular 7mm diametersThe side of the tabletting label of 9mm diameters covers, then use 3% cellulose acetate and 1.2% polyethylene glycol 2000 (with acetoneAnd formic acid is solvent, 9: 1V/V) solution coating.40 DEG C of tabletting is dried overnight, and the weightening of unidirectional releasing layer coating is 2mg.FinallySticky paper is peeled off, as unidirectional release channel.

10.4 enteric layers

The tabletting for not having L30D-55 in bio-adhesive layer uses enteric coating.Coating suspensions include 200g L30D-55,12g superfine talcum powders, 6g PEG6000, final volume 400ml in water.Coating is carried out in small-sized coating pan, and 40 DEG C of tabletting is dryDry 14hr.Enteric layer weightening 25mg.

Effect of 10.5 oral insulins to blood glucose

Using Beagle dogs evaluation two-layered coating (bio-adhesive of the label coating containing L30D-55 of infusion growth hormone release inhibiting hormoneLayer, is then coated unidirectional releasing layer) and three layers of coating (bio-adhesive layer of the label coating without L30D-55, then coating is unidirectionalReleasing layer, be then coated enteric layer) oral insulin tabletting effect.Experimentation is the same as embodiment 9.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.Growth hormone release inhibiting hormone (1 μ g/kg/min) is transfused by the ductus venosus of sacculus infusion pump, after being transfused 4h10ml water feeding tablettings.Feed is limited during experiment.Beasle dog experiment packet is shown in Table 19.

Table 19：Oral insulin experiment packet situation

Group	n	Administration
			Blank group	3	Comfort piece

Hypodermic injection group	3	3.5U/ dogs, it is dissolved in 0.01N HCl
			Without enteric coating	3	50U two-layered coating insulin tablettings
Enteric coating	4	Tri- layers of coating insulin tablettings of 50U

Different time takes blood, and blood glucose is tested using blood glucose meter and corresponding test paper.Figure is shown in influence of the oral insulin to blood glucose9。

As a result show, blood sugar level is significantly reduced after oral insulin tabletting, it is similar to injection group, and blank group is then firstBegin quickly to increase after reducing.There is no the tabletting group of enteric layer, 1h starts to reduce blood glucose after the tablet has been ingested, and the pressure containing enteric coatingPiece group then goes out an identical effect after 4h.

The effect of this result Exenatide similar with being formulated is consistent.The formula for illustrating two-layered coating is feasible in clinic's.

Embodiment 11

Dry method direct tablet compressing

Without granulation direct tablet compressing be simple economy tablet producing technology, can keep complete drug crystallization state andStability in technique.For low dosage tabletting, one be exactly the problem of main component during direct tablet compressing homogeneityProblem.

Using direct compression method, trial prepares insulin and Exenatide tabletting respectively.Component list is shown in Table 20.InsulinAnd Exenatide powder crosses 200 mesh sieves, auxiliary material is added, is well mixed.Using gearing tabletting mode, while detect the hard of tablettingDegree, friability and thickness.

Table 20：Piece core component (unit：mg)

According to USP<905>Test the homogeneity of insulin or Exenatide batch mixing.The powder of same piece weight is taken after mixing at randomMaterial, insulin or Exenatide content are surveyed using RP-HPLC.Mixed homogeneity is shown in Table 21.

Table 21：The homogeneity of direct compression method batch mixing

Batch	Medicine	Content/piece weight	Average value	RSD
					178-2	Exenatide	600μg/200mg	672±83	12.4%
176	Insulin	1mg(27U)/200mg	26±0.7	2.8%

As a result show, unit dose is that the batch mixing homogeneity of 1mg (account for piece weight 0.5%) insulin is to receive's.And 0.6mg Exenatide homogeneity is poor.The water imbibition of Exenatide powder may add more by force the difficulty of homogeneityDegree.

Therefore, although direct tablet compressing process is feasible, heterogeneity such as water imbibition and particle diameter of medicine etc. can influenceHomogeneity.

Embodiment 12

Non-solvent granulation in tabletting preparation process

Direct compression method problems faced be component homogeneous sex chromosome mosaicism, especially low dose of medicine, such as Ai SainaPeptide, and wet granulation can influence the stability of medicine.Therefore, employ a kind of non-solvent method of granulating and prepare insulin and Ai SaiThat peptide tabletting.The component list of different batches and different pharmaceutical is shown in Table 22~26.

Insulin and Exenatide powder cross 200 mesh sieves, are suspended to afterwards in 8% PVP ethanol solutions.Suspension is slightUltrasound is to without obvious bulky grain.In addition to superfine silica gel powder and magnesium stearate, all auxiliary materials are weighed, and are premixed in granulator.Medicine suspendsLiquid adds auxiliary material mixture, granulation, crosses 18 mesh sieves, vacuum drying afterwards.Superfine silica gel powder and magnesium stearate are added after drying, is usedInterlock tabletting.The hardness of tabletting, friability and thickness are detected simultaneously.

Table 22：Piece core component (the unit of the insulin containing 1mg：mg)

Table 23：Component (the unit of the label of the insulin containing 0.8mg：Mg, batch 182)

Table 24：Piece core component (the unit of the Exenatide containing 0.6mg：Mg, batch 178-1)

Table 25：Piece core component (the unit of the Exenatide containing 0.4mg：Mg, batch 183)

Table 26：Piece core component (unit containing 0.4mg Exenatides：Mg, batch 184)

Insulin or Exenatide content in tabletting are tested using RP-HPLC, according to USP<905>Evaluate the homogeneous of contentProperty.It the results are shown in Table 27：

Table 27：The content uniformity of insulin or Exenatide in 178~184 batch samples

Batch	Medicine	Content/piece weight	Average value	RSD
					178-1	Exenatide	600μg/220mg	583±6	1.0%
181	Insulin	1mg(27U)/300mg	28±0.2	0.8%
					182	Insulin	0.8mg(22U)/300mg	22±0.3	1.2%
183	Exenatide	400μg/300mg	424±14	3.3%
					184	Exenatide	400μg/300mg	393±9.4	2.4%

As a result show, the insulin or the content uniformity of Exenatide tabletting prepared using non-solvent method of granulating is canReceive.

Embodiment 13

Stability of the Exenatide in different formulations and preparation technology

The stability of Exenatide in different formulations is have rated, each formula is shown in Table 28.

Table 28：The stability of Exenatide

Formula	Exenatide	Technique
			Exenatide powder	1mg	N/A
Directly mixed with auxiliary material	1mg	N/A
			With auxiliary material tabletting	1mg	Wet granulation
With auxiliary material tabletting	1mg	Direct tablet compressing
			With auxiliary material tabletting	1mg	Non-solvent is pelletized
Add auxiliary material and gelatin, mannitol tabletting	1mg	Wet granulation
			Add auxiliary material and PEG tablettings	1mg	Non-solvent is pelletized

Auxiliary material used is listed as follows in basic recipe：

Table 29：Solvent (unit in label：mg)

Sample is placed in culture dish, is placed in varying environment and is continued 10 days, including 60 DEG C, 25 DEG C and 92.5% are relatively wetUnder degree, 4500lux illumination conditions, respectively at 0,5 and 10 day using RP-HPLC detection Exenatide contents.

The stability of different formulations is shown in Figure 10 A~10C.As a result show that Exenatide powder is relatively stable, in high temperature, high humidityAnd under illumination condition, it can still keep more than 95% within 10 days.Auxiliary material has similar stability after directly mixing, and shows in tablettingAuxiliary material and Exenatide are compatible.

However, the tabletting of wet granulation is highly unstable, the heavy losses at 5 days and 10 days.Add gelatin or mannitolDo not increase stability.When reducing the stability, especially high humility of Exenatide on the contrary, this is probably due to gelatinOr mannitol absorbs substantial amounts of moisture and caused under conditions of high humidity.

On the contrary, the tabletting pelletized using direct tablet compressing or non-solvent has preferable stability, with Ai SainaPeptide powder is similar.It is worth noting that, in non-solvent pelletization add low molecule amount PEG, stability still compared withDifference, this is probably to cause because Exenatide is partially dissolved in low molecule amount PEG.

Embodiment 14

Effect of the oral insulin to normal dogs

14.1 prepared by label

Matched according to table 30, being pelletized and being used using non-solvent interlocks tablet press machine compressed cores.Except insulin, PVP, micro mistOutside silica gel and magnesium stearate, remaining component is weighed and is well mixed in granulator in advance.Insulin powder is suspended in using 8%PVP ethanol solutions in, slight ultrasound.Suspension is added in auxiliary material and pelletized, is dried in vacuum overnight.Add superfine silica gel powderAnd magnesium stearate and it is well mixed after it is tabletted.The hardness of detection label, thickness, friability.

Table 30：Label matches (unit：mg)

14.2 bio-adhesive layer

Label uses 6%HPMC and 1.5%PEG6000 solution, is coated in coating pan (BY300A, Huanghai Sea instrument).StickLayer weightening 4mg.

14.3 unidirectional releasing layer

Tabletting is further coated using semi-permeable cellulose acetate.First will using the sticky paper of circular 7mm diametersThe side of the tabletting label of 10mm diameters covers, then molten using 3% cellulose acetate and 1.2% polyethylene glycol 2000 ethanolLiquid is coated.Tabletting drying at room temperature, the weightening of unidirectional releasing layer coating is 2mg.Finally sticky paper is peeled off, it is logical as unidirectional releaseRoad.

14.4 enteric layers

Tabletting is further coated enteric layer.Coating suspensions include 8%L30D-55,2%PEG6000.Coating is in small packageCarried out in clothing pot.Enteric layer weightening 20mg.

The blood sugar decreasing effect of 14.5 oral insulins

Using the effect of normal beasle dog evaluation oral insulin tabletting.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.Using the direct feeding tabletting of 10ml water.Feed is limited during experiment.Beasle dog experiment packetIt is shown in Table 31.

Table 31：Oral insulin experiment packet situation

Group	n	Administration
			Blank group	3	Comfort piece
Hypodermic injection group	3	3.5U/ dogs, it is dissolved in 0.01N HCl
			50U	3	2 × 25U oral insulin tablettings

Different time takes blood, and blood glucose is tested using blood glucose meter.Figure 11 is shown in influence of the oral insulin to blood glucose.

As a result prove that oral insulin tabletting significantly reduces blood sugar level, it is similar with injection of insulin, and blank group blood glucoseKeep constant.

The blood sugar level of normal individual is normally controlled in narrow range.In normal beasle dog, oral insulin tablettingThe blood glucose of induction reduces similar to the intensity and pattern of insulin injection.

Embodiment 15

Laser ablation process in tabletting preparation process

15.1 prepared by label

Matched according to table 32, using gearing tabletting compressed cores.In addition to Exenatide, superfine silica gel powder and magnesium stearate, itsRemaining component is weighed and is well mixed.Exenatide is suspended in 8% PVP ethanol solutions, as adhesive.After granulationVacuum drying.22 mesh sieves are crossed, add superfine silica gel powder and magnesium stearate, tabletting after being well mixed.Detect the hardness of label, thickness,Friability.

Table 32：Core tablet formula (unit：mg)

15.2 bio-adhesive layer

Label uses 3%HPMC E0 and 0.6%L30D-55 suspension, is wrapped in coating pan (BY300A, Huanghai Sea instrument)Clothing.Coating tablet is in 40 DEG C of dry 14h, adhesion layer weightening 3mg.

15.3 unidirectional releasing layer

Tabletting is further coated using semi-permeable cellulose acetate.When hand-manipulated, the tabletting label of 9mm diameters is firstFirst side is covered with the sticky paper of 7mm diameters of circle, then using 3% cellulose acetate and 1.2% polyethylene glycol 2000(solvent acetone and formic acid, 9: 1V/V) solution coating.Sticky paper is peeled off after drying, as unidirectional release channel.

During laser boring, tabletting is covered without sticky paper, but tabletting is integrally coated using cellulose acetate solution, after coating40 DEG C of tabletting be dried overnight.Go out the hole of 7mm diameters in wafer surface ablation using laser ablation apparatus (CMS) afterwards, withEffect when hand-manipulated is similar.Shown by the stability test of 2h in simulate the gastric juice, it is glutinous that laser boring will not destroy biologyThe integrality of attached layer.

In manual operations or laser drilling process, the weightening of unidirectional releasing layer is 2~3mg.

The absorption of 15.4 oral Exenatides

The effect of manual pasting or the unidirectional delivery formulations of oral Exenatide of laser boring is evaluated using normal beasle dog.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.Using the direct feeding tabletting of 10ml water.Feed is limited during experiment.Different time points useThe anticoagulant tube of test tube of hepari takes blood.Beasle dog experiment packet is shown in Table 33.

Table 33：Oral Exenatide experimental animal packet

Group	n	Administration
			Blank group	3	Comfort piece
Hypodermic injection group	3	60 μ g/ dogs, are dissolved in 10mM sodium acetate solutions, pH4.5
			Paster group	3	1mg, oral Exenatide tabletting, drilling after manual pasting
Laser boring group	3	1mg, oral Exenatide tabletting, laser boring

Different time points take blood, and serum is taken after 3000rpm centrifugations 10min, -20 DEG C of freezings of blood serum sample, is tried using ELISAAgent box (Phoenix) tests blood concentration.

Sample sets Exenatide plasma concentration curve is shown in Figure 12.Exenatide prepared by manual pasting drilling or laser boringThe drug absorption of unidirectional release tabletting is similar.

Embodiment 16

Insulin dose is transfused the effect of dog to growth hormone release inhibiting hormone

16.1 prepared by label

Matched according to table 34, using crawl tabletting compressed cores.In addition to insulin, superfine silica gel powder and magnesium stearate, remainingComponent is weighed and is well mixed.Pelletized using 8% PVP ethanol solutions (25%) as adhesive.It was dried in vacuo after granulationNight.Particle crosses 22 mesh sieves, according to the piece of every again needs, each weighs required grain amount, is then respectively adding insulin, micro-Powder silica gel and magnesium stearate, tabletting after being well mixed.The hardness of detection label, thickness, friability.

Table 34：Label matches (unit：mg)

16.2 bio-adhesive layer

16.3 unidirectional releasing layer

Tabletting is further coated using semi-permeable cellulose acetate.The tabletting label of 10mm diameters is first with circleThe sticky paper of 7mm diameters covers side, then using 3% cellulose acetate and the (solvent acetone of 1.2% polyethylene glycol 2000And formic acid, 9: 1V/V) solution coating.40 DEG C are dried overnight.Unidirectional releasing layer weightening 2mg, afterwards peels off sticky paper, as unidirectionalRelease channel.

Effect of 16.4 oral insulins to blood glucose

Using the effect of the beasle dog evaluation oral insulin of growth hormone release inhibiting hormone infusion.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.The direct feeding tabletting of 10ml water is used after infusion growth hormone release inhibiting hormone 4h.Limited during experiment intoFood.Beasle dog experiment packet is shown in Table 35.

Table 35：Oral insulin experimental animal is grouped

Group	n	Administration
			Blank group	3	Comfort piece
Injection group	3	Subcutaneous insulin injections, 3.5U/ dogs, molten and 0.01N HCl
			25U	3	25U tablettings, it is a piece of

25Ux2	4	25U tablettings, two panels
			50U	3	50U tablettings, it is a piece of

Different time points take blood, and blood glucose meter surveys blood glucose, and the blood sugar decreasing effect of oral insulin is shown in Table 13.

As a result show, oral insulin tabletting can significantly reduce blood glucose, similar to insulin injection.The oral pancreas islet of 25UFor element compared with the oral insulin of 50U or 25U × 2, blood glucose decline has dose dependent.The oral insulin of 50U or 25U × 2Blood glucose declines almost equivalent.

Embodiment 17

The influence that bio-adhesive polymer discharges and absorbed to Exenatide

17.1 prepared by label

Exenatide tabletting uses alcohol granulation, and method is the same.Proportioning is shown in Table 36.Exenatide powder is suspended in 8%PVP absolute ethyl alcohols in, ultrasound is to without obvious bulky grain.In addition to superfine silica gel powder and magnesium stearate, remaining component is weighed and madeIt is well mixed in grain machine.Exenatide suspension is added in auxiliary material mixture and pelletized, is dried in vacuo after crossing 18 mesh sieves.Add micro-Powder silica gel and magnesium stearate, tabletting after being well mixed.The hardness of detection label, thickness, friability.

Table 36：Core tablet formula (unit：mg)

17.2 bio-adhesive layer

Label is using 3%HPMC E50 and 0.6%L30D-55 water slurry (50% ethanol) in coating pan (BY300A, HuangExtra large instrument) in be coated.Or using 2.6%HPMC, 0.8%L30D-55 and 0.6%PEG suspension (50% ethanol) coating, bagGarment piece is in 40 DEG C of dry 14h, 3~4mg of adhesion layer weightening.

17.3 unidirectional releasing layer

Tabletting is further coated using semi-permeable cellulose acetate.Coating solution is 3.2% ethyl cellulose, 0.6%85% ethanol solution of polyethylene glycol 2000 and 0.2% triacetin.40 DEG C are dried overnight after coating, unidirectional releasing layer weightening2mg。

Tabletting uses laser boring, in the side of 9mm diameter tablettings cutting 7mm holes, as unidirectional release channel.

The absorption of 17.4 Exenatides

Using the assimilation effect of normal beasle dog evaluation Exenatide tabletting.

Healthy beasle dog 12,8~12kg of body weight, is responsible for nursing by animal center.Beasle dog is randomly divided into different controlTreatment group, overnight fasting before experiment.Using the direct feeding tabletting of 10ml water.Feed is limited during experiment.Beasle dog experiment packetIt is shown in Table 37.

Table 37：Oral Exenatide experiment packet

Group	n	Administration
			Blank group	3	Comfort piece
Hypodermic injection group	3	60 μ g/ dogs, are dissolved in 10mM sodium acetate solutions, pH4.5
			Exenatide group	3	0.4mg × 2 piece, 65%HPMC coatings
Exenatide group	3	0.4mg × 2 piece, 80%HPMC coatings

Different time points take blood, and serum is taken after 3000rpm centrifugations 10min, -20 DEG C of freezings of blood serum sample, is tried using ELISAAgent box (Phoenix) tests blood concentration.Sample sets Exenatide plasma concentration curve is shown in Figure 14.

As a result show that bio-adhesive layer is essential in tabletting, and the content of bio-adhesive polymer can significantly affectThe release of Exenatide and assimilation effect.