CN108283715A - A kind of analogue antigen compound is for treating the related indication purposes of HBV infection - Google Patents
A kind of analogue antigen compound is for treating the related indication purposes of HBV infection Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
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Abstract
The related indication method of HBV infection is treated the present invention relates to a kind of, the method includes being administered to the related indication subject of HBV infection comprising following two-part analogue antigen compound by therapeutically effective amount：(1)ε‑CH3(CH2)14CO (NH) KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA.The method of the present invention can induce HBV infection patients serum's HBeAb sun turn, HBeAg/HBeAb Virus mutations, reduce patients serum's HBV DNA levels, reduce Serum ALT levels, repair damage liver cell；Therefore, the related indication effective ways of HBV infection can be treated as a kind of.
Description
Technical field
The present invention relates to a kind of analogue antigen compounds for treating the related indication purposes of HBV infection.
Background technology
Chronic hepatitis B (abbreviation hepatitis B) is that one kind is caused by hepatitis type B virus (Hepatitis B virus, HBV)
, based on liver inflammatory lesion and a kind of disease of multiple organ injury can be caused.Hepatitis B is in worldwide prevalence, but differently
The epidemic strength in area is widely different.China belongs to the high Endemic Area of hepatitis B, and the HBsAg positive rates of population are 9.09%, are cured
Difficulty, poor prognosis seriously endanger health.Include at present mainly antiviral therapy for the treatment of hepatitis B, immune modulating treatment, resist
Inflammation treatment and antifibrosis therapy.It includes that interferons are similar with nucleosides (acid) to generally acknowledge effective Anti-HBV drugs both at home and abroad mainly
Object respectively has its advantage and disadvantage.The former the advantages of is that the course for the treatment of is relatively fixed, and HBeAg frequence of seroconversion is higher, and curative effect is relatively lasting, resistance to
Medicine variation is less；The disadvantage is that needing drug administration by injection, adverse reaction is more apparent, is unsuitable for liver function decompensation person.The latter's is excellent
Point is oral medication, inhibits virus function strong, and adverse reaction is few and slight, can be used for liver function decompensation person, the disadvantage is that treating
Journey is opposite to be not fixed, and HBeAg frequence of seroconversion is low, and curative effect is not lasting enough, and prolonged application can generate Resistance mutation, can go out after drug withdrawal
It is existing that sb.'s illness took a turn for the worse.Therefore, clinically there is an urgent need for one kind capable of preferably treating the related indication drug of HBV infection at present.
It is CH that a kind of primary structure is disclosed in CN1483736 (A) (be included in by reference herein)3(CH2)14The analogue antigen polypeptide of COKSSQYIKANSKFIGITEAAAFLPSDFFPSVGGGDPRVRGLYFPA, and disclose described more
Peptide can generate HBV neutrality antibodies in mouse Immune inducing in vivo, and induce and generate cell factor and cell-cytotoxic reaction.However, mesh
It is preceding that there is no the analogue antigen polypeptides to be used for the related indication report of clinical treatment HBV infection.
Invention content
The first aspect of the present invention provide it is a kind for the treatment of the related indication method of HBV infection, the method includes controlling
It treats and a effective amount of is administered to the related indication subject of HBV infection comprising following two-part analogue antigen compound：(1)
ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA, wherein treating
HBV infection related symptoms include improving HBeAg/HBeAb Virus mutations rate, improving the positive rate of HBeAb turns of serum, reduce serum
HBV DNA levels reduce Serum ALT levels and/or improve hepar damnification.
The second aspect of the present invention is provided is used to prepare raising comprising following two-part analogue antigen compound
The purposes of the drug of HBeAg/HBeAb Virus mutation rates：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and
(2)AAAFLPSDFFPSVGGGDPRVRGLYFPA。
The third aspect of the present invention is provided is used to prepare raising serum comprising following two-part analogue antigen compound
HBeAb turns the purposes of the drug of positive rate：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA。
The fourth aspect of the present invention is provided is used to prepare reduction serum comprising following two-part analogue antigen compound
The purposes of the drug of HBV DNA levels：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA。
The fifth aspect of the present invention is provided is used to prepare reduction serum comprising following two-part analogue antigen compound
The purposes of the drug of ALT levels：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA。
The sixth aspect of the present invention is provided is used to prepare improvement liver comprising following two-part analogue antigen compound
The purposes of the drug of damage：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA。
The method of the present invention can induce HBV infection patients serum's HBeAb sun turn, HBeAg/HBeAb Virus mutations, reduce
Patients serum's HBV DNA levels reduce Serum ALT levels, repair damage liver cell；Therefore, a kind of HBV that treats can be used as to feel
Contaminate related indication effective ways.
Description of the drawings
According to the detailed description carried out referring to the drawings, above and other aspects, features and advantages of the invention can become
It obtains clearer.
Fig. 1 shows that analogue antigen compound passes through the chromatography collection of illustrative plates of reversed-phased high performace liquid chromatographic after purification.
Fig. 2 shows the analogue antigen compounds-prepared in the embodiment of the present invention 1 observed by transmission electron microscope (TEM)
The analogue antigen prepared in the ultra microstructure (Fig. 2 a) of liposome and the embodiment of the present invention 1 detected by laser particle size analyzer
The particle diameter distribution (Fig. 2 b) of drug-lipid body.
Fig. 3 show Chronic Hepatitis B receive analogue antigen compound treatment before (Fig. 3 a) and treat after (Fig. 3 b) liver
Dirty tissue morphology (HE is dyed, × 200).
Specific implementation mode
As described above, treating the related indication method of HBV infection the present invention provides a kind of, the method includes treating
It is a effective amount of to be administered to the related indication subject of HBV infection comprising following two-part analogue antigen compound：(1)ε-
CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA, wherein treating
HBV infection related symptoms include improving HBeAg/HBeAb Virus mutations rate, improving the positive rate of HBeAb turns of serum, reduce serum
HBV DNA levels reduce Serum ALT levels and/or improve hepar damnification.
Term as used herein " therapeutically effective amount " refers to the amount with advantageous effect in treated subject, i.e.,
Be enough to cure, mitigate or part prevent given disease and its complication clinical manifestation amount.
In an embodiment of the method for the present invention, the HBV infection related symptoms are one or more of disease
Symptom：Chronic hepatitis B, hepatic sclerosis and hepatocellular carcinoma.
In the method for the invention, the subject refers to the mankind and non-human mammal, such as, but not limited to inhuman
Primate, cat, dog, sheep, goat, horse, ox, pig and rodent (such as, but not limited to mouse and rat)；And it is non-
Mammal, such as, but not limited to birds, poultry, reptile, amphibian.The subject can be arbitrary gender, appoint
What at age.Term " subject " and " patient " are used interchangeably herein.In a preferred embodiment, described tested
Person is the mankind.
In an embodiment of the method for the present invention, the CH in the part (1) of the analogue antigen compound3(CH2)14CO- is by CH3(CH2)16CO- is replaced.
In an embodiment of the method for the present invention, AAA in the part (2) of the analogue antigen compound and/or
GGG is replaced by SSS.
In another embodiment of the method for the present invention, the part (1) and part (2) of the analogue antigen compound are logical
Cross covalent linkage.
In a specific embodiment of the method for the present invention, the analogue antigen compound is by liposome with shape
At analogue antigen drug-lipid body.In a specific embodiment, the analogue antigen drug-lipid body it is straight
Diameter is 80-100nm.In another specific embodiment, the chemical combination-composite lipidosome also includes pharmaceutically acceptable assistant
Agent, carrier or auxiliary material.In one embodiment, the analogue antigen drug-lipid body is pharmaceutically acceptable arbitrary
Dosage form.In a preferred embodiment, the analogue antigen drug-lipid body be liquid dosage form or freeze-dried formulation, more
Preferably freeze-dried formulation.
In an embodiment of the method for the present invention, the preparation of the analogue antigen compound be injection, transdermal agent,
Oral agents, inhalant or suppository.In a preferred embodiment, the preparation of the analogue antigen compound is injection.
In the method for the invention, the analogue antigen compound can be administered with any administration route, it is described to give
Medicine approach include but not limited to oral cavity, rectum, nose, lung, Epidural cavity, eye, ear, intra-arterial, in heart, in skin, intravenous, flesh
Meat is interior, in intraperitoneal, bone, in bladder, it is subcutaneous, part, percutaneous and given through mucous membrane, such as by sublingual, oral cavity, vagina and sucking
Medicine approach.
In a preferred embodiment in accordance with this invention, using the analogue antigen compound as injection with subcutaneous way
Diameter is administered.In a specific embodiment, the analogue antigen compound is administered by upper limb or abdominal part hypodermic
To adult patient.In another specific embodiment, the analogue antigen compound is subcutaneously injected by anterolateral thigh
It is administered to child patient.
In a specific embodiment, given with 45 degree of depth of needle using the analogue antigen compound as injection
Medicine to subject subcutaneous fat.In a specific embodiment, the analogue antigen compound is given as injection
Injection point when medicine is to subject is 1-3, and is administered 3-21 times, is preferably administered 6-21 times.
In one embodiment of the invention, the analogue antigen compound is administered in two stages, wherein
6 times (first 4 times per minor tick 3-4 week, the 5th, 6 time per minor ticks 7-8 weeks) are administered in first stage, and second stage is administered 15 times (preceding 9
It is secondary per minor tick 3 weeks, latter 6 times every minor tick 4 weeks).In embodiments of the invention, institute is administered with the dosage of 300-1800 μ g
State analogue antigen compound.In a preferred embodiment, the analogue antigen is administered with the dosage of 600 or 900 μ g
Close object.In a further preferred embodiment, the analogue antigen compound is administered with the dosage of 900 μ g.
The present invention also provides be used to prepare treatment HBV infection correlation disease comprising following two-part analogue antigen compound
The purposes of the drug of shape：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA, wherein the treatment HBV infection related symptoms include improving HBeAg/HBeAb blood
It is clear to learn conversion ratio, improve the positive rate of HBeAb turns of serum, reduce serum HBV DNA level and/or reduce Serum ALT levels.
In one embodiment, the HBV infection related symptoms are the symptom of one or more of disease：Chronic
Type hepatitis, hepatic sclerosis and hepatocellular carcinoma.
In one embodiment, the CH in the part (1) of the analogue antigen compound3(CH2)14CO- is by CH3
(CH2)16CO- is replaced.In another embodiment, AAA the and/or GGG quilts in the part (2) of the analogue antigen compound
SSS is replaced.In another embodiment, the part (1) of the analogue antigen compound and part (2) pass through covalent bond chain
It connects.
In a specific embodiment, compound-that the compound is configured to be formed by liposome
Liposome.In a specific embodiment, a diameter of 80-100nm of the analogue antigen drug-lipid body.One
In a embodiment, the analogue antigen drug-lipid body is pharmaceutically acceptable arbitrary dosage form.It is preferred real at one
It applies in scheme, the analogue antigen drug-lipid body is liquid dosage form or freeze-dried formulation, more preferably freeze-dried formulation.
In one embodiment, the drug also includes pharmaceutically acceptable adjuvant, carrier or auxiliary material.At another
In embodiment, the drug is injection, transdermal agent, oral agents, inhalant or suppository.In a preferred embodiment
In, the drug is injection.
In one embodiment, the drug is excellent for the mankind and non-human mammal and non-mammalian animal
It is selected to the mankind.
In embodiments of the invention, the drug can be administered with any administration route, the administration route packet
Include but be not limited to oral cavity, rectum, nose, lung, Epidural cavity, eye, ear, intra-arterial, in heart, in skin, intravenous, intramuscular, abdomen
In intracavitary, bone, in bladder, it is subcutaneous, part, percutaneous and through mucous membrane, such as pass through sublingual, oral cavity, vagina and inhalation approach.
In a preferred embodiment in accordance with this invention, using the drug as injection with subdermal routes of administration.
In one specific embodiment, the drug is administered to adult patient by upper limb or abdominal part hypodermic.At another
In specific embodiment, the drug is passed through into anterolateral thigh subcutaneous administrations to child patient.
In a specific embodiment, subject is administered to 45 degree of depth of needle using the drug as injection
Subcutaneous fat.In a specific embodiment, injection point when subject is administered to using the drug as injection
It is 1-3, and is administered 3-21 times, is preferably administered 6-21 times.
In one embodiment of the invention, the drug is administered in two stages, the wherein first stage is given
Medicine 6 times (first 4 times per minor tick 3-4 week, the 5th, 6 time per minor tick 7-8 week), 15 times (first 9 times per minor ticks for second stage administration
3 weeks, latter 6 times per minor tick 4 weeks).In embodiments of the invention, the drug is administered with the dosage of 300-1800 μ g.
In one preferred embodiment, the drug is administered with the dosage of 600 or 900 μ g.In a further preferred embodiment,
The drug is administered with the dosage of 900 μ g.
Illustrate present disclosure below by way of specific embodiment.It should be understood that the specific embodiment is only to illustrate mesh
, it is not meant to that present disclosure is only limitted to specific embodiment.The analogue antigen compound or reagent used in embodiment
It can be bought by commercial sources, or be prepared by conventional method well known by persons skilled in the art；Used experiment
Instrument can be bought by commercial sources.
Embodiment 1：The synthesis and purifying of analogue antigen compound
1) synthesis of analogue antigen compound
The sequence of the analogue antigen compound of the present invention is ε-CH3(CH2)14CO-(NH)-
KSSQYIKANSKFIGITEAAAFLPSDFFPSVG GGDPRVRGLYFPA are synthesized using Fmoc solid-phase synthesis.
Fixed c-terminus extends peptide chain from C-terminal to N-terminal.It is enterprising in Applied Biosystems 431A Peptide synthesizers
Row.Fmoc amino acid is raw material, and applied sample amount 1mM, side chain protection is：Ser (tBu), Thr (tBu), Tyr (tBu), His (Trt),
Gln(Trt)、ASP(OtBu)、Glu(OtBu)、Arg(Pmc).Selection HMP- resins are solid phase carrier, and applied sample amount 0.25mM is former
Material/resin is than 4：1.The amino acid (first amino acid of polypeptide c-terminus) being coupled on resin is lived using symmetric anhydride activation method
Change, the activation of remaining amino acid and palmitic acid uses HOBt/DCC activation methods.Wherein, asparagine, arginine and palmitic acid are adopted
With double couple crosslinking, nothing takes off Fmoc blocking group steps after palmitic acid coupling.
2) cracking of analogue antigen compound-resin
Select TFA lysates (0.75g phenol, 0.25mL dithioglycols, 0.5mL thioanisoles, 0.5mL deionized waters,
10.0mL TFA) it is cracked, the generation of side reaction can be inhibited to the maximum extent.It is first 1.5mL, reaction in lysate volume
Under conditions of temperature is 25 DEG C and the reaction time is 2.0 hours, analogue antigen compound-resin concentration is measured to cracking reaction point
Influence from effect, as a result as shown in table 1 below：
Influence of 1. analogue antigen compound of the table-resin concentration to cracking Reaction Separation effect
※Note：Reflect the change of " yield " indirectly with " analogue antigen compound/analogue antigen compound-resin ((%) "
Change.
From table 1 it follows that as analogue antigen compound-resin concentration excessively high (53.33mg/mL), lysate adds
Precipitation can be generated by entering DMSO, and the analogue antigen compound of separation accounts for 26.03 ﹪ of analogue antigen compound-resin, and yield is relatively low；
When analogue antigen compound-resin concentration is 40mg/mL, lysate is added DMSO and does not generate precipitation, the analogue antigen of separation
Compound accounts for 29.80 ﹪ of analogue antigen compound-resin, and yield is higher.Therefore, by analogue antigen chemical combination in cracking reaction
The concentration of object-resin is determined as 40mg/mL.
Then influence of the cracking reaction time to cracking Reaction Separation effect is further measured, as a result as shown in table 2 below：
The influence that the 2 cracking reaction time of table isolates and purifies analogue antigen compound-resin lysate
※Note：Reflect the variation of " yield " indirectly with " analogue antigen compound/analogue antigen compound-resin (%) ".
From Table 2, it can be seen that by cracking reaction time control in 1-2h, after cracking, the analogue antigen that finally obtains
The homozygosis and yield of compound are higher, therefore the cracking reaction time is determined as 1.5 hours.
3) purifying of analogue antigen compound
The lysate containing analogue antigen compound is obtained under the crack reacting condition of above-mentioned determination, then uses two steps
Method purifies analogue antigen compound：Size exclusion chromatography method is used to carry out preliminary purification, tomographic system first：P-6000
Pump and AKTAexplorer 100；Chromatographic column：Diameter 10mm, column length 250mm, filler Sephadex LH-20；Column temperature：25℃；
Mobile phase：DMSO；Flow velocity：0.4ml/min.Collect analogue antigen compound component then use reversed-phased high performace liquid chromatographic into
Row purifying, tomographic system：50 R1 columns of AP-5 50/275POROS and AKTA explorer 100；Chromatographic column：Diameter 50mm,
50 R1 of column length 275mm, filler POROS；Column temperature：34℃；Mobile phase A：30% ethyl alcohol -10mmol/L phosphoric acid, Mobile phase B：
90% ethyl alcohol -10mmol/L phosphoric acid；Flow velocity：100ml/min.Fig. 1 shows that analogue antigen compound passes through RP-HPLC
The chromatography collection of illustrative plates of chromatography after purification.After measured, by above-mentioned two-step purifying, the purity of analogue antigen compound reaches 98.66
± 0.14%, yield is 86.89 ± 0.43%, has reached preferable refining effect.
Embodiment 2：The preparation of analogue antigen drug-lipid body
1) analogue antigen drug-lipid body is prepared using second emulsifying method：Lipid composition is dissolved in diethyl ether solution, then
The concentrate of the analogue antigen compound obtained with embodiment 1 mixes, and forms emulsion (W/O), injects phosphate buffer
(PB) or in water, while temperature and stirring are controlled, secondary formation emulsion (W/O/W) gradually forms mould with the volatilization of ether
Quasi- antigen compound-liposome.Pass through ultrafiltration apparatus and dialysis (dialysis multiple must reach 200 times or more) and 10 μm of micropore filters
Membrane filtration is obtained with removing analogue antigen drug-lipid body may dissociating in liquid and that aggregate and precipitate may occur
Analogue antigen drug-lipid body concentrate.Then it is sweet 20% will to be added in 0.5L analogue antigen drug-lipid body concentrates
Reveal alcohol solution 140mL, 20% human serum albumin 30mL, 30mL phosphate buffer, after mixing well, by 1mL/ bottles of specification
It is dispensed.After E-C Apparatus company Supermodulyo types freeze dryers are cooled to 8 DEG C in advance, above-mentioned sample is put into freeze-drying
Machine is cooled to -39 DEG C overnight；Condenser, vacuum pump and heater are opened successively, organic solvent and moisture in the sample that distils,
Finally obtain white loose block；It sterilizes and is packaged to be finished product.
2) after the analogue antigen drug-lipid body of preparation freeze-drying finished product being resuspended with 1mL aseptic injection waters, with 0.22 μm
Millipore polycarbonate membrane filters be filtered to remove impurity, the analogue antigen drug-lipid liquid solution of 100 μ L is taken, with nothing
Water-ethanol is diluted to 2mL, and analogue antigen drug-lipid liquid solution is resuspended, is sufficiently stirred；With liquid-transfering gun by the sample drop of preparation
In on support film (copper mesh), dry；Transmission electron microscope is opened, the copper mesh for being loaded with sample is put into transmission electron microscope and is observed, is put
Big multiple is respectively × 160000, as a result as shown in Figure 2 a.From Fig. 2 a as can be seen that analogue antigen drug-lipid body at
Single vesica exists, and size is relatively uniform, most of to be less than 100nm.
3) the analogue antigen drug-lipid body grain for using the detection of Malvern ZEN1690 type laser particle size analyzers to prepare
Diameter size.After the analogue antigen drug-lipid body of preparation freeze-drying finished product is resuspended with 1mL aseptic injection waters first, with 0.22 μ
The polycarbonate membrane filter of m is filtered to remove impurity, the analogue antigen drug-lipid liquid solution of 100 μ L is taken, with the PBS of 0.01M
(pH7.4) it is diluted to 2mL, is sufficiently stirred, 1.2mL is taken to be added in sample container, opens computer program, sets detection parameters, in advance
Heat engine device 30min, addition sample are detected, as a result as shown in Figure 2 b.As can be seen that laser particle analyzer analysis knot from Fig. 2 b
Fruit is similar with Electronic Speculum result, and liposome size distribution is 30-250nm, and 70-95% or more liposomal diameters are in 80-100nm
In range.
Embodiment 3：Therapeutic effect of the analogue antigen drug-lipid body in Chronic Hepatitis B
The present embodiment utilizes the analogue antigen drug-lipid body finished product prepared in embodiment 1, to the chronic hepatitis B of screening
Patient treats, and studies its therapeutic effect and effect in chronic hepatitis B disease.
(1) selection of study subject：
First stage (0-76 weeks) selects HBeAg positive chronics hepatitis B patient 360 as treatment object.It is divided into
3 groups below, every group of 120 subjects：One group is given 600 μ g analogue antigen drug-lipids body+300 μ g empty liposomes (600 μ
G treatment groups)；One group is given 900 μ g analogue antigen drug-lipids bodies (900 μ g treatment groups)；One group is given 900 μ g sky lipids
Body (control group).
Second stage (76-144 weeks)：In three groups of subjects of above-mentioned completion research in 76 weeks, if having virology after treatment
Response but serum-free response/have serology response but virus-free response/virology serology is unresponsive, but be interested in continuing with
The subject for participating in research has 183 people (wherein to include 58 people of 600 μ g treatment groups of first stage, 900 μ g treatment groups of first stage
60 people and first stage control group 65 people), continue to give the treatment of 900 μ g analogue antigen drug-lipid bodies, and follow-up is seen
It examines to 3 years (144 weeks).
(2) administering mode：
First stage：It is administered using upper arm or abdominal part hypodermic mode, 45 ° of inserting needles, depth and subcutaneous fat, injection point are
1-3.600 μ g treatment groups give 600 μ g analogue antigen drug-lipids body+300 μ g empty liposomes；900 μ g treatment groups are given
900 μ g analogue antigen drug-lipid bodies；Control group gives 900 μ g empty liposomes.Administration time is respectively the 0th, 4,8,12,
20,28 weeks.
Second stage：Respectively the 80th, 83,86,89,92,95,98,101,104,108,112,116,120,124 and
128 weeks, 900 μ g analogue antigen drug-lipid bodies, injection site and mode same first stage, co-injection 15 times is subcutaneously injected.
(3) curative effect evaluation：
It observes in (0-76 weeks) in the first stage and second stage (76-144 weeks), detection is made by the embodiment of the present invention 1
After standby analogue antigen drug-lipid body treatment, the anti-HBe frequence of seroconversion of HBeAg/, the anti-HBe of patient turn positive rate, serum
HBV DNA copy numbers and serum glutamic pyruvic transminase (ALT) are horizontal.Specifically, in the first stage treat after, share 354 it is tested
Person is included into Intentionality treatment (ITT) crowd and (is defined as：It is all through randomized grouping, at least used once studied medicine
The subject of curative effect index (the anti-HBe of HBeAg/) data after object and at least once medication) carry out following results statistics；
Wherein, control group 119,1 people because after no medication curative effect achievement data be removed；600 μ g treatment groups 119,1 people is because of nothing
Curative effect achievement data is removed after medication；900 μ g treatment groups 116,4 people are because of curative effect achievement data quilt after no medication
It rejects.After second stage treatment,
3 people of control group, 600 μ g treatment groups, 3 people, 900 μ g treatment groups, 2 people because after no medication curative effect achievement data picked
It removes；The Intentionality treatment number for being finally included in statistics is respectively 62 people of control group, 600 μ g treatment groups, 55 people and 900 μ g treatment groups
58 people amount to 175 people.
1. after the treatment of analogue antigen drug-lipid body, the higher anti-HBe Virus mutations rates of HBeAg/ occur for patient：
For HBeAg positive chronic hepatitis B patients, HBeAg Virus mutations, that is, HBeAg disappears, anti-Hbe turns
For the positive.The anti-HBe Virus mutations of HBeAg/ occur, is satisfied with terminal as treating chronic hepatitis B, indicates long-term prognosis
Improve, slows down with progression of disease as hepatic sclerosis incidence is reduced.At the end of studying in the first stage, the HBeAg/ of control group is anti-
HBe Virus mutation rates are 20.2% (24/119).235 subjects of whole analogue antigen drug-lipid body drug
In (ITT crowd), there are 79 subjects that the anti-HBe Virus mutations of HBeAg/, conversion ratio 33.6% occurs；Wherein, 600 μ g are controlled
Treatment group and 900 μ g treatment groups Virus mutation rates are respectively 28.6% (34/119) and 38.8% (45/116) (table 3).
In second stage treatment end, Virus mutation rate when entering compared to 76 weeks group, the patient of former each group is again
After receiving the treatment of 900 μ g analogue antigen drug-lipid bodies, Virus mutation rate has a more substantial increase.Wherein control group
Occur Virus mutation subject's ratio from 76 weeks when 17.7% (11/62) promotion to 144 weeks end points when 38.7%
(24/62)；600 μ g treatment groups from 76 weeks when 20.0% (11/55) promoted to 144 weeks end points when 40.0% (22/55),
900 μ g treatment groups from 76 weeks when 29.3% (17/58) promoted to 144 weeks end points when 34.5% (20/58) (table 4).
The 1st stage of table 3. (0-76 weeks), the anti-HBe Virus mutations rates of HBeAg/
The 2nd stage of table 4. (76-144 weeks), the anti-HBe Virus mutations rates of HBeAg/
According to the data (table 5) in hepatopathy association of U.S. chronic hepatitis B practice guideline in 2016, receive the present invention's
The anti-HBe Virus mutations rates of HBeAg/ are better than nucleoside analogue drugs (A Defu after the treatment of analogue antigen drug-lipid body
Wei, Lamivudine and Entecavir) and long-acting interferon.
The anti-HBe Virus mutations rate * of HBeAg/ after the treatment of 5. clinical treatment hepatitis B key agents of table
* data are derived from " AASLD Guidelines for Treatment of Chronic Hepatitis B ",
Hepatology.2016；63(1):261-83.
2. after the treatment of analogue antigen drug-lipid body, patient occurs to turn positive rate compared with highly resistance HBe：
HBe antibody turns sun, i.e. serum HBe antibody occurs.HBe antibody can reflect host immune response situation；HBe antibody turns
The immune response that more stable HBV specificity is obtained in sun prompt host, is conducive to the control of HBV infection.First
At the end of stage is studied, it is 29.4% (35/119), whole analogue antigen drug-lipid body that the anti-HBe of control group, which turns positive rate,
In 235 subjects (ITT crowd) of drug, there are 95 subjects that anti-HBe occurs and turn sun, conversion ratio 40.4%.600 μ g are controlled
It is respectively 37.0% (44/119) and 44.0% (51/116) (table 6) that the anti-HBe for the treatment of group and 900 μ g treatment groups, which turns positive rate,.
In second stage treatment end (table 7), Virus mutation rate when entering compared to 76 weeks group, control group and 600 μ
After the patient of g treatment groups receives the treatment of 900 μ g analogue antigen drug-lipid bodies again, it is equal that anti-HBe turns positive Virus mutation rate
It is improved.Wherein, control group occur subject's ratio of Virus mutation from 76 weeks when 35.3% (22/62) promotion to 144
41.9% (26/62) when all end points；600 μ g treatment groups from 76 weeks when 27.3% (15/55) promoted to 144 weeks end points
When 47.3% (26/55).
The 1st stage of table 6. (0-76 weeks), anti-HBe turn the ratio of sun
The 2nd stage of table 7. (76-144 weeks), the ratio that anti-HBe sun turns
3. after the treatment of analogue antigen drug-lipid body, subject's peripheral blood HBV DNA copy numbers reduce：
Peripheral blood HBV DNA are formed by being discharged into blood by the liver cell of HBV infection, are reflection HBV infection treatments
The most direct index of effect.The reduction of peripheral blood HBV DNA copy numbers then means that hbv replication is suppressed, and is reflected and is controlled
The validity for the treatment of means.At the end of studying in the first stage, the subject of control group, 600 μ g treatment groups and 900 μ g treatment groups sends out
Raw HBV DNA carrying capacity declines the virological response rate of >=2 Logarithmic degrees, respectively 40.3% (48/119), 48.7% (58/
And 50.0% (58/116) (table 8) 119).
In second stage treatment end, when control group the 76th week serum HBV DNA (IU/ml) carrying capacity decline >=2 it is right
The ratio of the patient of several levels is 27.4% (17/62), reaches 59.7% (37/62) within the 144th week.When 600 μ g treatment groups the 76th week
Serum HBV DNA (IU/ml) carrying capacity decline more than or equal to 2 Logarithmic degrees Proportion of patients be 27.3% (15/55), the 144th
Reach 56.4% (31/55) week.Serum HBV DNA (IU/ml) carrying capacity, which declines, when 900 μ g treatment groups the 76th week is greater than or equal to 2
The Proportion of patients of a Logarithmic degree is 37.9% (22/58), reaches 51.7% (30/58) (table 9) within the 144th week.
The 1st stage of table 8. (0-76 weeks), serum HBV DNA decline the percentage of >=2 Logarithmic degrees
The 2nd stage of table 9. (76-144 weeks), serum HBV DNA decline the percentage of >=2 Logarithmic degrees
4. after the treatment of analogue antigen drug-lipid body, the Serum ALT levels of subject reduce：
ALT is generated by liver cell specificity, is normally inside liver cell.When liver cell is damaged, then largely
ALT is significantly increased by entering peripheral blood, Serum ALT inside liver cell.Therefore, Serum ALT levels reflect hepatocellular injury feelings
Condition：Serum ALT levels rise, and prompt the generation of hepar damnification；The decline of Serum ALT levels then prompts effective control of HBV infection
System, the reduction of hepar damnification.At the end of studying in the first stage, control group, 600 μ g treatment groups and 900 μ g treatment groups it is tested
Person's ALT levels are down to the ratio of normal range (NR), respectively 34.5% (41/119), 37.8% (45/119) and 48.3% (56/
116) (table 10).
The 1st stage of table 10. (0-76 weeks), ALT levels are down to the ratio of normal range (NR)
5. after the treatment of analogue antigen drug-lipid body, the hepatic pathology of subject is improved：
Hepatic pathology section is to reflect the most intuitive evidence of hepar damnification situation.The same Chronic Hepatitis B is chosen,
It is pretherapy and post-treatment in analogue antigen drug-lipid body, liver organization is obtained in a manner of liver puncture, observes pretherapy and post-treatment liver
Pathological change situation.Treatment results as shown in figure 3, by two stages treatment, liver oedema, balloon sample become degree obviously subtract
Gently；Point, focal necrosis significantly reduce；Lymphocytic infiltration significantly reduces.It can be effectively improved slowly it can be seen that the drug has
Property hepatitis B patient hepatic pathology, reduce hepar damnification, be conducive to hepatitis B infected effective control.
To sum up, present invention demonstrates that the analogue antigen drug-lipid body can effectively induce HBV infection patients serum
HBeAb sun turn, HBeAg/HBeAb Virus mutations reduce patients serum's HBV DNA levels, reduce Serum ALT levels, repair
Damage liver cell；Therefore, it can be used for preparing and treat the above-mentioned related indication drug of HBV infection.
Claims (10)
1. being used to prepare the drug for improving HBeAg/HBeAb Virus mutation rates comprising following two-part analogue antigen compound
Purposes：(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2)
AAAFLPSDFFPSVGGGDPRVRGLYFPA。
2. being used to prepare the purposes for improving the drug that serum HBeAb turns positive rate comprising following two-part analogue antigen compound：
(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA.
3. being used to prepare the purposes for the drug for reducing serum HBV DNA levels comprising following two-part analogue antigen compound：
(1)ε-CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA.
4. being used to prepare the purposes for the drug for reducing Serum ALT levels comprising following two-part analogue antigen compound：(1)ε-
CH3(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA.
5. being used to prepare the purposes for the drug for improving hepar damnification comprising following two-part analogue antigen compound：(1)ε-CH3
(CH2)14CO- (NH)-KSSQYIKANSKFIGITE and (2) AAAFLPSDFFPSVGGGDPRVRGLYFPA.
6. the purposes of any one of claim 1-5, wherein the CH in the part (1) of the analogue antigen compound3(CH2)14CO
By CH3(CH2)16CO is replaced, and the AAA sequences and/or GGG sequences in the part (2) of the analogue antigen compound are replaced by SSS
It changes, or the part (1) of the wherein described analogue antigen compound and part (2) pass through covalent linkage.
7. the purposes of any one of claim 1-5, wherein the analogue antigen compound is configured to by liposome shape
At analogue antigen drug-lipid body；Preferably, a diameter of 80-100nm of the analogue antigen drug-lipid body；Or
The wherein described analogue antigen drug-lipid body of person is pharmaceutically acceptable arbitrary dosage form；Preferably, the analogue antigen
It is liquid dosage form or freeze-dried formulation, more preferably freeze-dried formulation to close object-liposome.
8. the purposes of any one of claim 1-5, wherein the drug also includes pharmaceutically acceptable adjuvant, carrier or auxiliary
Material；Preferably, the drug is injection, transdermal agent, oral agents, inhalant or suppository；It is highly preferred that the drug is injection
Agent.
9. the purposes of any one of claim 1-5, wherein the drug is administered with any administration route, the administration way
Diameter include but not limited to oral cavity, rectum, nose, lung, Epidural cavity, eye, ear, intra-arterial, in heart, in skin, intravenous, muscle
In interior, intraperitoneal, bone, in bladder, it is subcutaneous, part, percutaneous and through mucous membrane, such as pass through sublingual, oral cavity, vagina and inhalation
Approach；Preferably, using the drug as injection with subdermal routes of administration；It is highly preferred that by the drug by upper limb or
Abdominal part hypodermic is administered to adult patient or the drug is passed through anterolateral thigh subcutaneous administrations to child patient；And
The subcutaneous fat of subject is wherein administered to 45 degree of depth of needle using the drug as injection；Preferably, by the medicine
Injection point when object is administered to subject as injection is 1-3, and is administered 3-21 times, is preferably administered 6-21 times.
10. the purposes of any one of claim 1-5, wherein the drug is administered in two stages, in the first stage
Administration 6 times, wherein first 4 times per minor tick 3-4 week, the 5th, 6 every minor tick 7-8 week；It is administered 15 times in second stage, wherein before
9 times per minor tick 3 weeks, latter 6 times every minor tick 4 weeks；And wherein the drug is administered with the dosage of 300-1800 μ g
The drug；Preferably, the drug is administered the drug with the dosage of 600 or 900 μ g；It is highly preferred that will be described
Drug is administered with the dosage of 900 μ g.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711384035.XA CN108283715A (en) | 2017-12-20 | 2017-12-20 | A kind of analogue antigen compound is for treating the related indication purposes of HBV infection |
| PCT/CN2018/120491 WO2019120112A1 (en) | 2017-12-20 | 2018-12-12 | Use of mimetic antigen compound for treating symptoms related to hbv infections |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711384035.XA CN108283715A (en) | 2017-12-20 | 2017-12-20 | A kind of analogue antigen compound is for treating the related indication purposes of HBV infection |
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| CN107028887A (en) * | 2017-03-28 | 2017-08-11 | 江苏孟德尔基因科技有限公司 | Liposome is used for the purposes for treating chronic viral hepatitis type B |
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| WO2019120112A9 (en) | 2020-02-13 |
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