CN103212075B - A kind of eye drops containing VEGF antagonist - Google Patents
A kind of eye drops containing VEGF antagonist Download PDFInfo
- Publication number
- CN103212075B CN103212075B CN201210017896.5A CN201210017896A CN103212075B CN 103212075 B CN103212075 B CN 103212075B CN 201210017896 A CN201210017896 A CN 201210017896A CN 103212075 B CN103212075 B CN 103212075B
- Authority
- CN
- China
- Prior art keywords
- eye drops
- compaq
- cornea
- western general
- eye
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- 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
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/12—Carboxylic acids; Salts or anhydrides thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0048—Eye, e.g. artificial tears
Abstract
The present invention relates to a kind of eye drops containing VEGF antagonist, the VEGF antagonist containing low concentration in the eye drops, especially containing the VEGF antagonist of 0.05 9.0mg/ml, its is evident in efficacy on the basis of safety and stability is ensured for eye drops of the present invention.
Description
Technical field
The present invention relates to field of medicine preparations, and in particular to a kind of eye drops containing VEGF antagonist.
Background technology
The ocular such as cornea rebirth blood vessel, neovascular glaucoma, pteryium, chronic conjunctivitis disease morbidity with newly
The overexpression for having certain relation, VEGF of angiogenic can induce ocular new vessels to be formed.Wherein cornea rebirth
Blood vessel is not a kind of independent keratonosus, but a kind of pathological change.The reason for causing cornea rebirth blood vessel is a lot, immune inflammation
Property, infectivity, denaturation, it is traumatic, contact lenses it is improper use, iatrogenic disease etc..Cornea rebirth caused by above-mentioned reason
If blood vessel is treated not in time, corneal blindness is eventually resulted in.Cornea rebirth blood vessel is the master of world wide visual loss and blindness
Reason is wanted, the ophthalmic patient in the U.S. about 4.14% suffers from cornea rebirth blood vessel, the U.S. there are 1,400,000 newly-increased cornea rebirth blood every year
Pipe patient, wherein 12% meeting causes visual impairment, has the cornea rebirth blood vessel of about 170,000 newly-increased visual impairments every year in the U.S.
Patient.The Chinese still not epidemiology survey on cornea rebirth blood vessel, if accounting for ophthalmology according to U.S.'s cornea rebirth blood vessel
Patient 4%, wherein 12% can cause visual impairment to calculate, national ophthalmic patient is 62,830,000 within 2009, and the country there are about every year
300000 newly-increased patient affects one's power of vision because of cornea rebirth.Patient's cornea rebirth blood vessel enters exhibition causes the further evil of eyesight
Change, finally need corneal transplantation to recover eyesight.Therefore such patient needs to carry out effective anti-new vessels treatment, in case
Only eyesight is further lost, while avoiding carrying out corneal transplantation, doctor does not have effective treatment means for cornea rebirth blood vessel.
Bio-pharmaceutical, especially recombinates albuminoid numerous characteristics and is totally different from small-molecule chemical molecule, and it occurs unstable
It is a multi-step reaction to determine degradation reaction.It is limited to the limitation of present analysis method, these for betiding higher structure are full of many
The degraded of individual step also is difficult to be accurately measured, its stability, and preservation especially under normal conditions is exactly one and very big chooses
War.Eyes are one of most important organs of human body：The information of nearly the 80% of brain comes from eyes；Eyes are special due to its simultaneously
Physiological make-up, is the weak tissue of comparing, and to viscosity, the requirement such as osmotic pressure is all very high.Therefore for eye-drops preparations, especially
There is the eye-drops preparations requirement used when wound abnormal strict.
The biological effect of VEGF (vascular endothelial growth factor) is by its specific receptor VEGFR (blood vessels
Endothelial cell growth factor receptor 2 body) come what is realized, VEGFR can cause by ligand-mediated Dimerized, the dimerization of acceptor for mediation
Body promotes adjacent receptor subunit autophosphorylation and dephosphorylation, so that trigger signal is transduceed.VEGFR is tyrosine protein
Kinases, is divided into by its function and structure：Fins- samples EGFR-TK -1 (VEGFR-1/Flt-1), kinase insert domain receptor
(VEGFR-2/Flk-1/KDR), fins- samples EGFR-TK -4 (VEGFR-3/Flt-4) and some low relative molecular masses
VEGFR(neuropilin-1).Flt-1 and KDR are distributed mainly on vascular endothelial cell, and Flt-4 is then distributed mainly on lymph
On endothelial cell.Predominantly Flt-1 and KDR, the Flt-1 being combined with VEGF are high compared with KDR with the adhesion of VEGF, Ta Mendou
It is glycosylated transmembrane receptor, directly participates in VEGF and enter intracellular signal transmission.Flt-1 can promote blood after being combined with VEGF
The formation of endothelial cell and regulation vascular permeability；KDR can then promote after being combined with VEGF vascular endothelial cell hyperplasia and
It is ripe.VEGF antagonist, especially the target gene engineered protein of a class engineer, such as Lucentis, Avastin, VEGF-
Trap etc., they can effectively block the signal transmission mediated by VEGF (VEGF), suppress lesion newborn
The growth of blood vessel, and for treating the disease that tumour, eyes etc. are caused by new vessels, but the system for having listed of said medicine
Dosage form formula be injection, its administering mode be vitreum direct injection or Formulations for systemic administration, its dosage form for patients, its
Compliance is poor, and there is inconvenience.Therefore, a kind of researcher's patient compliance that begins one's study is good, and containing easily and effectively
The pharmaceutical dosage forms of VEGF antagonist, US7303748 discloses a kind of eye drops containing VEGF-trap, its preparation prescription：
39.4-103.06mg/ml VEGF-trap, 5mM phosphoric acid, 5mM citric acids, 100mM sodium chloride, 0.005% Tween-20.But
The eye drops concentration is larger, because biological agent is usually present unstable problem, the Anti-angiogenic therapy patient of current ocular
Many to be all reluctant hospitalization, the biological agent of high concentration easily causes the inconvenience for preserving and using, while ocular administration is
One special method of administration, to pH, osmotic pressure and local irritation, it is impossible to add bacteriostatic agent and antiseptic etc., above mentioned problem
So that for the costly of the anti-new vessels medicine of ocular, effect is limited.Therefore it provides a kind of concentration is low, suppress new
The eye drops of angiogenic determined curative effect, and use it for treatment cornea rebirth blood vessel, neovascular glaucoma, it is pteryium,
Various eye surface diseases such as chronic conjunctivitis, have significant meaning for meeting clinical demand.
The content of the invention
But the present invention needs to solve that one of technical problem is to provide a kind of small determined curative effect of concentration contains VEGF antagonisms
The eye drops of agent.
In order to solve the above-mentioned technical problem, the invention provides following technical scheme：
One aspect of the present invention provides a kind of eye drops containing VEGF antagonist, and 0.05- is contained in the eye drops
The VEGF antagonist of 9.0mg/ml；Preferably comprise the VEGF antagonist of 0.1-1.0mg/ml；Further preferably 0.5mg/ml's
VEGF antagonist；Wherein described VEGF antagonist is preferably the fusion protein of the segment containing FLT-1 and KDR.
Above-mentioned VEGF antagonist preferably has the fusion protein of one of following structure：
A.FP1, is immunized by the 2nd immunoglobulin-like region of FLT-1 and the 3rd immunoglobulin-like region of KDR with people
The albumen of immunoglobulin Fc fragment fusion：FLTd2-KDRd3-Fc；
B.FP2, by the 1st immunoglobulin-like region of KDR, the 2nd immunoglobulin-like region of FLT-1 and the 3rd of KDR the
Immunoglobulin-like region and the albumen of human immunoglobulin(HIg) Fc segment compositions：
KDRd1-FLTd2-KDRd3-Fc；
C.FP3, is exempted from by the 2nd immunoglobulin-like region of FLT-1 and the 3-4 immunoglobulin-likes region of KDR with people
The albumen of epidemic disease immunoglobulin Fc fragment fusion：FLTd2-KDRd3,4-Fc；
D.FP4, by the 2nd immunoglobulin-like region of FLT-1, the 3rd immunoglobulin-like region of KDR and FLT-1's
4th immunoglobulin-like region and the albumen of human immunoglobulin(HIg) Fc segment compositions：
FLTd2-KDRd3-FLTd4-Fc；
E.FP5, is exempted from by the 2nd immunoglobulin-like region of FLT-1 and the 3-5 immunoglobulin-likes region of KDR with people
The albumen of epidemic disease immunoglobulin Fc fragment fusion：FLTd2-KDRd3,4,5-Fc；Or
F.FP6, by the 2nd immunoglobulin-like region of FLT-1, the 3rd immunoglobulin-like region of KDR and FLT-1's
4-5 immunoglobulin-likes region and the albumen of human immunoglobulin(HIg) Fc segment compositions：
FLTd2-KDRd3-FLTd4,5-Fc；
G.FP7, by the 3-4 immunoglobulin-likes region fusion in the 2nd immunoglobulin-like region and KDR of FLT-1
Into albumen：FLTd2-KDRd3,4；
H.FP8, is formed by the 3rd immunoglobulin-like region fusion in the 2nd immunoglobulin-like region and KDR of FLT-1
Albumen：FLTd2-KDRd3.
The amino acid FLT-1D2 in above FLT-1 and KDR immunoglobulin-like region, FLT-1D4, KDRD1, KDRD3,
KDRD4 is shown in that the amino acid sequence of sequence table 1-5, FP3 albumen is shown in that sequence table 7 is shown in sequence table 6, the amino acid sequence of FP1 albumen,
The amino acid sequence of FP7 albumen is shown in sequence table 8, and the amino acid sequence of FP8 albumen is shown in sequence table 9.
In the eye drops that the present invention is provided, can also one or more containing following components：
A () 5-100mM buffer solutions, acid therein is selected from Tris-HCl, citric acid, phosphoric acid, dibastic sodium phosphate, biphosphate
Sodium, acetic acid, succinic acid, one or more in hydrochloric acid；
B () 5-500mM basic amino acids are selected from lysine, one kind or its combination in arginine, and histidine；
C () 0.1-30% salt penetrations press agent conditioning agent, sugar therein to be selected from sucrose, trehalose, mannitol, glycerine, the third two
Alcohol, one or more in sorb ester alcohol, salt is selected from sodium chloride or other one kind or its group pharmaceutically in acceptable salt
Close；
One or more surfactant or cosolvent of (d) 0.005-0.1%, selected from polyethylene glycol, polysorbas20, tween
80, propane diols, dimethyl sulfoxide (DMSO) or other pharmaceutically in acceptable surfactant one or more；
The pH value of solution that one or more of said components are made into is 6~8.3.
VEGF antagonist described in above-mentioned eye drops is most preferably such as SEQ ID No：6 fusion protein.
The present invention still further provides the eye drops containing following component, and concrete component is as follows：
(a) 0.05-9mg/ml such as SEQ ID No：6 fusion protein；
The citric acid of (b) 5-250mM；
The arginine of (c) 5-500mM or histidine one or two；
The sucrose or trehalose of (d) 4-30%；
The surfactant or cosolvent of (e) 0.01-0.1%, one or two selected from polyethylene glycol or polysorbas20；
F () regulation pH is 7.5~8.3.
The present invention further provides the eye drops containing following component, and concrete component is as follows：
(a) 0.1-1mg/ml such as SEQ ID No：6 fusion protein；
The citric acid of (b) 10-50mM；
The arginine of (c) 50-100mM or histidine one or two；
The sucrose of (d) 5-20%；
The surfactant or cosolvent of (e) 0.01-0.1%, one or two selected from polyethylene glycol or polysorbas20；
F () regulation pH is 7.5~8.3.
Invention still further provides the eye drops containing following component, concrete component is as follows：
(a) 0.5mg/ml such as SEQ ID No：6 fusion protein；
The citric acid of (b) 50mM；
The arginine of (c) 250mM；
The sucrose of (d) 12.5%；
The Tween-20 of (e) 0.05%；
F () regulation pH is 7.5~8.3.
The medicine of the eye surface diseases that treatment is caused by angiogenesis or growth is being prepared present invention also offers above-mentioned eye drops
Purposes in thing；Preferably eye surface diseases are the postoperative new vessels of corneal transplantation, cornea rebirth blood vessel, ocular new vessels or the wing
Any one or its complication of the shape triangular mass of mucous membrane growing from the inner corner of the eye.
Technology contents are used as the application's disclosed in Chinese patent ZL200510073595.4 and ZL200610066257.2
With reference to；The technology contents of the undisclosed patent CN 201010267503.7 of China also serve as the reference of the application.
The present invention compared with prior art it is advantageous that：The content of fusion protein is extremely low in eye drops and determined curative effect,
Production cost can be significantly saved on the basis of low concentration eye drops stability higher.
Specific embodiment
Following examples to of the invention only as being explained further, it is impossible to used as being limit to the scope of the present invention
System.
The western general eye drops of embodiment 1, Compaq causes cornea rebirth blood vessel research to alkali burn
Medicine：Aureomycin hydrochloride eye ointment, specification lot number：2.0g/ branch, 411002, valid until 2014.12, Chongqing section is auspicious
Pharmaceutical Co；Lidocaine hydrochloride injection, specification lot number：5ml/ branch, 0.1g/ branch, valid until 2012.04.,
Medicine Jiaozhuo, Tianjin Co., Ltd.
Reagent：NaOH (NaOH), specification lot number：500g/ bottles, 20091223, Chengdu Ke Long chemical reagents factory.
Given the test agent：
A, western general eye drops (being prepared according to the embodiment 3) 10mg/ml of Compaq, colourless transparent liquid, 1ml/ branch, lot number：
FR1108001, is stored in 2~8 DEG C, and ocular surface is added dropwise to when using；
B, western general eye drops (being prepared according to the embodiment 3) 9mg/ml of Compaq, colourless transparent liquid, 1ml/ branch, lot number：
FR1108002, is stored in 2~8 DEG C, and ocular surface is added dropwise to when using；
C, dexamethasone, colourless transparent liquid, 1ml/ branch, lot number：FR1108003, is stored in 2~8 DEG C, is added dropwise when using
Enter ocular surface；
D, western general eye drops (being prepared according to the embodiment 3) 0.1mg/ml of Compaq, colourless transparent liquid, 1ml/ branch, lot number：
FR1108004, is stored in 2~8 DEG C, and ocular surface is added dropwise to when using；
E, without the western general Formulation Buffer (buffer) (being prepared according to embodiment 16) of Compaq, colourless transparent liquid, 1ml/
Branch, lot number：FR1108005, is stored in 2~8 DEG C, and ocular surface is added dropwise to when using；
F, western general eye drops (being prepared according to the embodiment 3) 5mg/ml of Compaq, colourless transparent liquid, 1ml/ branch, lot number：
FR1108006, is stored in 2~8 DEG C, and ocular surface is added dropwise to when using；
There is provided by chemical drug research department Drug Manufacturing Room in Kang Hong medicine companies conglomerate technique center.
Experimental technique and result：
Health is taken without eye diseases new zealand rabbit 48, claims its body weight, (1ml/kg) is anaesthetized with 3% yellow Jackets, and in
Ocular surface bestows local anaesthetics lidocaine hydrochloride liquid, and dosage is 20 μ l/；9mm diameter filter papers are prepared, 1mol/ is soaked in
, be positioned over for filter paper with tweezers unnecessary NaOH solution sucked on dry filter paper by about 10s in LNaOH solution, will be soaked with
The filter paper of NaOH is removed after being placed in rabbit eyes cornea center 60s, and wash bottle about 20ml normal saline flushings angle is taken rapidly
Film, and give prevention antibiotic prevent infect (aureomycin hydrochloride eye ointment), 3 times/2 days.
It is divided into burn, A (western general eye drops (being prepared according to the embodiment 3) 10mg/ml of Compaq), B (Compaq west after modeling immediately
General eye drops (being prepared according to embodiment 3) 9mg/ml), C (dexamethasone), D ((according to embodiment 3 make by the western general eye drops of Compaq
It is standby) 0.1mg/ml), E (be free of the western general formulation buffer solution (being prepared according to embodiment 16) of Compaq) and F (the western general eye drips of Compaq
Liquid (being prepared according to embodiment 3) 5mg/ml) seven groups, the modeling same day is 0 day, and since the 1st day, A~F groups gave corresponding medicine
Thing, 6 times/day of administration frequency, dosage 50 μ l/, successive administration 10 days every time separately take 2 new zealand rabbits and are set to normal group.
Simultaneously, whether observation cornea NV growth conditions and eye have inflammatory reaction for administration daily.
In administration the 10th day, with 3% Nembutal sodium solution anesthetized animal (1ml/kg), and local anaesthesia is bestowed in ocular surface
Medicine lidocaine hydrochloride liquid, dosage is 20 μ l/, in 10 times of thing Microscopic observation rabbit cornea NV hour directions of slit-lamp, while
Taken a picture under 10 times and 16 times of object lens.Collection image carries out hour number correction in Photoshop CS, and cornea rebirth area is used
Image Pro Plus treatment；Area formula：S=C/12 × 3.1416 × [R2-(R-L)2], C represents the edge of cornea in picture
From having NV to without the hour number shared by NV growth time points, R represents the edge that is contacted with sclera from cornea in picture in cornea
The length of the heart, L represented from the root of cornea and sclera engagement edge NV to the end NV length of NV in cornea in picture, each
A length of vessel most long is taken in hour.All data statistic analysis are analyzed using T variance tests.Data withRepresent, be
Average value, s is standard deviation.
By general ocular it has been observed that the 1st, 2 day each group burn cornea marginal vessel net of burn is congested substantially；Burn
3rd day, edge of cornea had NV to grow；The 5th day western general eye drops 5mg/ml of Compaq and without the western general formulation buffer solution of Compaq
NV growths in cornea surface are obvious, the western general eye drops 10mg/ml of dexamethasone, Compaq, the western general eye drops 9mg/ml of Compaq and health
Bai Xipu eye drops 0.1mg/ml groups NV growths are not obvious；7th day each group cornea surface NV growth is obvious, burn group and is free of
The western general formulation buffer solution group cornea NV of Compaq grows to burn cornea, and part NV has grown into burn edge of cornea, and ground
The western general eye drops 10mg/ml of Sai meter Song, Compaq, the western general eye drops 9mg/ml of Compaq, the western general eye drops 5mg/ml of Compaq and Compaq
The western general few NV of eye drops 0.1mg/ml groups grow into burn edge of cornea；Burn group and western general without Compaq at the 10th day
The existing NV of formulation buffer solution group is grown into burn cornea, and the western general 10mg/ml of dexamethasone, Compaq, the western general eye drops of Compaq
It is less that the western general 5mg/ml of 9mg/ml, Compaq and the western general 0.1mg/ml groups NV of Compaq grow into burn cornea.
Cornea rebirth blood vessel area effect：10th day cornea NV area datas analysis result understand, with alkali burn group and not
The western general formulation buffer solution group containing Compaq is compared, the western general eye drops 9mg/ml of the western general 10mg/ml of dexamethasone, Compaq, Compaq,
The western general 5mg/ml of Compaq and the western general 0.1mg/ml of Compaq can substantially suppress cornea NV growths, NV areas be reduced, with statistics
It is optimal (P ＜ 0.01) that meaning (P ＜ 0.05), wherein dexamethasone general 0.1mg/ml western with Compaq suppress NV growths.Although being free of
The western general formulation buffer solution unrestraint cornea rebirth blood vessel growth of Compaq, and NV areas are more than burn group, but without significantly
Sex differernce.Result such as table 1.
Each medicine of table 1 causes the influence of rabbit cornea NV to alkali burn
Note：Negative control is the buffer solution of not drug containing, and positive control is dexamethasone, is compared with burn group,*P ＜
0.05,**P ＜ 0.01；Compare with B groups,#P ＜ 0.05,##P ＜ 0.01；Compare with E groups, ^P ＜ 0.05, ^^P ＜ 0.01.
The western general eye drops of the low dosage Compaq of embodiment 2 causes cornea NV growth effects to alkali burn
Medicine：Aureomycin hydrochloride eye ointment, specification lot number：2.0g/ branch, 411002, valid until 2014.12, Chongqing section is auspicious
Pharmaceutical Co；Lidocaine hydrochloride injection, specification lot number：5ml/ branch, 0.1g/ branch, valid until 2012.04.,
Medicine Jiaozhuo, Tianjin Co., Ltd.
Reagent：NaOH (NaOH), specification lot number：500g/ bottles, 20091223, Chengdu Ke Long chemical reagents factory.
Given the test agent：
A, western general eye drops (being prepared according to the embodiment 3) 0.5mg/ml of Compaq, colourless transparent liquid, 800 μ l/ branch, lot number：
20111001,2~8 DEG C are stored in, ocular surface is added dropwise to when using；
B, western general eye drops (being prepared according to the embodiment 3) 0.1mg/ml of Compaq, colourless transparent liquid, 800 μ l/ branch, lot number：
20111001,2~8 DEG C are stored in, ocular surface is added dropwise to when using；
C, without the western general formulation buffer solution of Compaq, colourless transparent liquid, 800 μ l/ branch, lot number：20111001, preserve
In 2~8 DEG C, ocular surface is added dropwise to when using；
D, western general eye drops (being prepared according to the embodiment 3) 0.01mg/ml of Compaq, colourless transparent liquid, 800 μ l/ branch, batch
Number：20111001,2~8 DEG C are stored in, ocular surface is added dropwise to when using；
E, dexamethasone, colourless transparent liquid, 800 μ l/ branch, lot number：20111001,2~8 DEG C are stored in, dripped when using
Add ocular surface；
F, western general eye drops (being prepared according to the embodiment 3) 0.05mg/ml of Compaq, colourless transparent liquid, 800 μ l/ branch, batch
Number：20111001,2~8 DEG C are stored in, ocular surface is added dropwise to when using；
G, western general eye drops (being prepared according to the embodiment 3) 1mg/ml of Compaq, colourless transparent liquid, 800 μ l/ branch, lot number：
20111001,2~8 DEG C are stored in, ocular surface is added dropwise to when using；
There is provided by chemical drug research department Drug Manufacturing Room in Kang Hong medicine companies conglomerate technique center.
Experimental technique and result：
Experiment is divided into A (western general eye drops (being prepared according to the embodiment 3) 0.5mg/ml of Compaq), B (the western general eye drops of Compaq
(being prepared according to embodiment 3) 0.1mg/ml), C (be free of the western general Formulation Buffer (being prepared according to embodiment 16) of Compaq
Buffer), D (western general eye drops (being prepared according to the embodiment 3) 0.01mg/ml of Compaq), E (dexamethasone), F (the western general drops of Compaq
Ocular fluid (being prepared according to embodiment 3) 0.05mg/ml) and G (western general eye drops (being prepared according to the embodiment 3) 1mg/ml of Compaq) eight
Group.
Test method is operated with embodiment 1.
By ocular it has been observed that the 1st, 2 day each group burn cornea marginal vessel net of burn is congested substantially, part eyelid
Red and swollen phenomenon；Burn the 3rd day, edge of cornea there are NV growth tendencies, eyelid is red and swollen to disappear；5th day, each group anterior corneal surface had NV to give birth to
It is long, but there is part lagophthalmos eyelid to occur red and swollen phenomenon again；Each group cornea surface NV growths in 7th day are obvious, there is significantly centripetal
Property growth, and Dexamethasone group cornea NV is less, but lagophthalmos eyelid redness phenomenon is more serious；It is 10th day, western general containing Compaq
Each group cornea NV growths are obvious, and part cornea rebirth blood vessel has grown into burned part, and Dexamethasone group NV grows into Corneal Burn
Position is less, and lagophthalmos eyelid redness phenomenon has no recovery, and does not have lagophthalmos eyelid redness phenomenon containing the western general each group of Compaq.
Cornea rebirth blood vessel area effect：Cornea NV area datas analysis result understands within 10th day, western general with without Compaq
Formulation buffer solution group compare, the western general eye drops 0.5mg/ml of dexamethasone, Compaq can substantially suppress cornea NV growth, subtract
Few NV areas, with statistical significance (P ＜ 0.05).The western general eye drops 1mg/ml of Compaq, the western general eye drops 0.1mg/ml of Compaq
Also cornea NV growths can be suppressed.And the western general eye drops 0.01mg/ml unrestraints cornea rebirth blood vessel growth of Compaq, and NV faces
Product is more than burn group, but there was no significant difference.It is shown in Table 2.
Each medicine of table 2 causes the influence of rabbit cornea NV to alkali burn
Note：Buffer is compareed,*P ＜ 005,**P ＜ 001.
The preparation of western general (FP3 fusion proteins) eye drops of embodiment 3, Compaq
Prescription：
FP3 fusion proteins 10mg/ml, 9mg/ml, 5mg/ml, 1mg/ml, 0.5mg/ml, 0.1mg/ml, 0.05mg/ml or
0.01mg/ml
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, contain 55mM citric acids, 12.5% sucrose, 250mM arginine and 0.05% by aseptic manipulation addition filtration sterilization
The buffer solution of polysorbas20, filtration, regulation FP3 fusion proteins are to 10mg/ml, 9mg/ml, 5mg/ml, 1mg/ml, 0.5mg/ respectively
Ml, 0.1mg/ml, 0.05mg/ml or 0.01mg/ml, pH are 7.5~8.3, aseptic subpackaged in the container for containing eye drops, are deposited
It is put in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of embodiment 4, Compaq
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, add being told containing 10mM citric acids, 8.0% sucrose, 5mM arginine and 0.05% for filtration sterilization by aseptic manipulation
The buffer solution of temperature 20, filtration, regulation FP3 fusion proteins to 0.5mg/ml, pH are 7.5~8.3, aseptic subpackaged to splendid attire eye drops
Container in, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 5
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, by aseptic manipulation add filtration sterilization containing 100mM citric acids, 20.0% sucrose, 250mM arginine and
The buffer solution of 0.10% polysorbas20, filtration, regulation FP3 fusion proteins to 1.0mg/ml, pH is 7.5~8.3 aseptic subpackaged to Sheng
Fill in the container of eye drops, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 6
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, by aseptic manipulation add filtration sterilization containing 5mM sodium dihydrogen phosphates, 10.0% trehalose, 100mM arginine and
The buffer solution of 0.01%PEG400, filtration, regulation FP3 fusion proteins to 0.1mg/ml, pH is 7.5~8.3, aseptic subpackaged to Sheng
Fill in the container of eye drops, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 7
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, contain 5mM citric acids, 4.0% sucrose, 4.0% sodium chloride, 100mM essence ammonia by aseptic manipulation addition filtration sterilization
Acid, 100mM histidines, 0.05% polysorbas20, the buffer solution of 0.05%PEG400, filtration, regulation FP3 fusion proteins to
0.05mg/ml, pH are 7.5~8.3, aseptic subpackaged in the container for containing eye drops, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 8
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, add being told containing 10mM citric acids, 30.0% sucrose, 500mM arginine, 0.1% for filtration sterilization by aseptic manipulation
The buffer solution of temperature 20, filtration, regulation FP3 fusion proteins to 5.0mg/ml, pH are 7.5~8.3, aseptic subpackaged to splendid attire eye drops
Container in, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 9
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, add being told containing 10mM citric acids, 5.0% sucrose, 100mM arginine, 0.05% for filtration sterilization by aseptic manipulation
The buffer solution of temperature 20, filtration, regulation FP3 fusion proteins to 0.5mg/ml, pH are 7.5~8.3, aseptic subpackaged to splendid attire eye drops
Container in, deposit in 2-8 DEG C.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 10
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, contain 250mM citric acids, 8.0% sucrose, 100mM histidines, 0.10% by aseptic manipulation addition filtration sterilization
The buffer solution of polysorbas20, filtration, regulation FP3 fusion proteins to 0.5mg/ml, pH are 7.5~8.3, aseptic subpackaged to splendid attire eye drip
In the container of liquid, 2-8 DEG C is deposited in.
The preparation of western general (FP3 fusion proteins) eye drops of the Compaq of embodiment 13
Prescription：
Preparation method：To concentrate after the FP3 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, add being told containing 10mM citric acids, 5.0% sucrose, 100mM arginine, 0.05% for filtration sterilization by aseptic manipulation
The buffer solution of temperature 20, filtration, regulation FP3 fusion proteins to 0.5mg/ml, pH are 7.5~8.3, aseptic subpackaged to splendid attire eye drops
Container in, deposit in 2-8 DEG C.
The preparation of the serial fusion protein eye drops of embodiment 14
Prescription：
Preparation method：Various fusion proteins (FP1, FP2, FP4, FP5, FP6, the FP7, FP8) stoste changed after liquid will be concentrated
After thawing, in clean bench (aseptic cabinet) is removed in C grades of cleaning, by aseptic manipulation add filtration sterilization containing 55mM citric acids,
12.5% sucrose, the buffer solution of 250mM arginine and 0.05% polysorbas20, filtering, respectively adjust fusion protein to 9mg/ml,
5mg/ml, 1mg/ml, 0.5mg/ml, 0.1mg/ml or 0.05mg/ml, pH are 6.0~8.3, aseptic subpackaged to splendid attire eye drops
Container in, deposit in 2-8 DEG C.
The preparation of embodiment 15FP1 fusion protein eye drops
Prescription：
Preparation method：To concentrate after the FP1 fusion protein stostes changed after liquid thaw, and go clean bench (aseptic in C grades of cleaning
Cabinet) in, add being told containing 5mM phosphoric acid, 5mM citric acids, 100mM sodium chloride and 0.005% for filtration sterilization by aseptic manipulation
The buffer solution of temperature 20, filtration, to 1.0mg/ml, pH is 6.0-8.0 to regulation fusion protein, aseptic subpackaged to the appearance for containing eye drops
In device, 2-8 DEG C is deposited in.
It is prepared by Formulation Buffer of the embodiment 16 without Compaq western general (FP3 fusion proteins)
Prescription：
Preparation method：55mM citric acids, 12.5% sucrose, 250mM will be contained by aseptic manipulation addition filtration sterilization
The buffer solution of arginine and 0.05% polysorbas20, filtration, respectively regulation pH be 7.5~8.3, it is aseptic subpackaged to containing eye drops
In container, 2-8 DEG C is deposited in.
Western general (FP3 fusion proteins) stability of the eye drops at 25 DEG C of the 0.1mg/ml Compaqs of embodiment 17
1st, western general (FP3 fusion proteins) eye drops of 0.1mg/ml Compaqs is prepared according to embodiment 3
2nd, by western general (FP3 fusion proteins) eye drops of above-mentioned 0.1mg/ml Compaqs, 25 DEG C keep sample, 0,1,2,3,4,5,6,
7,8,9,10,11, December determination sample determines stability by SEC-HPLC.Result shows, can effectively suppress to be polymerized at this
The generation of thing, product purity does not decline substantially, and fusion protein is almost unchanged with the affinity of VEGF.
Concrete outcome is shown in Table 3.
Stability of the table 3.0.1mg/ml FP3 fusion proteins at 25 DEG C
| Time (moon) | Outward appearance | PH value | Polymer (%) | Affinity (pM) |
| 0 | It is qualified | 7.2 | 0.01 | 10.6 |
| 1 | It is qualified | 7.4 | 0.03 | 10.5 |
| 2 | It is qualified | 7.5 | 0.03 | 10.4 |
| 3 | It is qualified | 7.5 | 0.05 | 10.4 |
| 6 | It is qualified | 7.7 | 0.07 | 10.2 |
| 9 | It is qualified | 7.6 | 0.08 | 10.3 |
| 12 | It is qualified | 7.5 | 0.09 | 10.1 |
Western general (FP3 fusion proteins) stability of the eye drops at 4 DEG C of the 10mg/ml Compaqs of embodiment 18
1st, western general (FP3 fusion proteins) eye drops of 10mg/ml Compaqs is prepared according to embodiment 3
2nd, by western general (FP3 fusion proteins) eye drops of above-mentioned 10mg/ml Compaqs, 4 DEG C keep sample, 0,1,2,3,4,5,6,7,
8,9,10,11, December determination sample determines stability by SEC-HPLC.Result shown, polymer can be effectively suppressed at this
Generation, polymer increases slow in product, and fusion protein is almost unchanged with the affinity of VEGF.Concrete outcome is shown in Table 4.
Stability of the table 4.10mg/ml FP3 fusion proteins at 4 DEG C
Claims (4)
1. a kind of eye drops containing VEGF antagonist, it is characterised in that contain
(a) 0.1-1mg/ml such as SEQ ID No:6 fusion protein；
The citric acid of (b) 10-50mM；
The arginine of (c) 50-100mM or histidine one or two；
The sucrose of (d) 5-20%；
The surfactant or cosolvent of (e) 0.01-0.1%, one or two selected from polyethylene glycol or polysorbas20；
F () regulation pH is 7.5~8.3.
2. eye drops according to claim 1, it is characterised in that contain
(a) 0.5mg/ml such as SEQ ID No:6 fusion protein；
The citric acid of (b) 10mM；
The arginine of (c) 100mM；
The sucrose of (d) 5%；
The Tween-20 of (e) 0.05%；
F () regulation pH is 7.5~8.3.
3. the eye drops according to any one of claim 1-2 is preparing the ocular that treatment is caused by angiogenesis or growth
Purposes in the medicine of disease.
4. eye drops according to claim 3 is preparing the medicine of the eye surface diseases that treatment is caused by angiogenesis or growth
In purposes, it is characterised in that the eye surface diseases be the postoperative new vessels of corneal transplantation, cornea rebirth blood vessel, the new green blood of ocular
Pipe or any one pteryium or its complication.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710357021.2A CN107115294B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
| CN201210017896.5A CN103212075B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210017896.5A CN103212075B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710357021.2A Division CN107115294B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103212075A CN103212075A (en) | 2013-07-24 |
| CN103212075B true CN103212075B (en) | 2017-06-27 |
Family
ID=48810455
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710357021.2A Active CN107115294B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
| CN201210017896.5A Active CN103212075B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710357021.2A Active CN107115294B (en) | 2012-01-19 | 2012-01-19 | A kind of eye drops containing VEGF antagonist |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN107115294B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11806398B2 (en) | 2005-03-25 | 2023-11-07 | Regeneron Pharmaceuticals, Inc. | Citrate buffered VEGF antagonist formulations |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3753548A1 (en) | 2006-06-16 | 2020-12-23 | Regeneron Pharmaceuticals, Inc. | Vegf antagonist formulations suitable for intravitreal administration |
| SG10201509193UA (en) | 2011-01-13 | 2015-12-30 | Regeneron Pharma | Use of a vegf antagonist to treat angiogenic eye disorders |
| CN107115294B (en) * | 2012-01-19 | 2019-10-18 | 北京康弘生物医药有限公司 | A kind of eye drops containing VEGF antagonist |
| CN104940926B (en) | 2014-09-25 | 2017-09-22 | 信达生物制药(苏州)有限公司 | Recombination fusion protein preparation |
| CN105435222B (en) | 2014-09-25 | 2018-05-29 | 信达生物制药(苏州)有限公司 | Recombination fusion protein preparation |
| KR20240043821A (en) | 2015-06-06 | 2024-04-03 | 클라우드브레이크 테라퓨틱스, 엘엘씨 | Compositions and methods for treating pterygium |
| EP3463315B1 (en) | 2016-06-02 | 2023-06-14 | ADS Therapeutics LLC | Compositions and methods of using nintedanib for treating ocular diseases with abnormal neovascularization |
| CN106890193A (en) * | 2017-04-21 | 2017-06-27 | 云南农业大学 | A kind of dog, cat stem cell eye-drops preparations and its application |
| CN116897052A (en) * | 2021-03-04 | 2023-10-17 | 百奥泰生物制药股份有限公司 | anti-VEGF antibody formulations |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101478949A (en) * | 2006-06-16 | 2009-07-08 | 瑞泽恩制药公司 | VEGF antagonist formulations suitable for intravitreal administration |
| CN102233132A (en) * | 2010-04-28 | 2011-11-09 | 成都康弘生物科技有限公司 | Application of VEGF acceptor fusion proteins in preparation of drugs for inhibiting growth of ocular surface neovascularization |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100567325C (en) * | 2006-03-31 | 2009-12-09 | 成都康弘生物科技有限公司 | Vegf receptor fusion rotein and the application in the medicine of preparation treatment disease of eye thereof |
| CN102380096B (en) * | 2010-08-31 | 2014-04-30 | 成都康弘生物科技有限公司 | Medicine combination containing fusion protein for suppressing angiogenesis and application |
| CN107115294B (en) * | 2012-01-19 | 2019-10-18 | 北京康弘生物医药有限公司 | A kind of eye drops containing VEGF antagonist |
-
2012
- 2012-01-19 CN CN201710357021.2A patent/CN107115294B/en active Active
- 2012-01-19 CN CN201210017896.5A patent/CN103212075B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101478949A (en) * | 2006-06-16 | 2009-07-08 | 瑞泽恩制药公司 | VEGF antagonist formulations suitable for intravitreal administration |
| CN102233132A (en) * | 2010-04-28 | 2011-11-09 | 成都康弘生物科技有限公司 | Application of VEGF acceptor fusion proteins in preparation of drugs for inhibiting growth of ocular surface neovascularization |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11806398B2 (en) | 2005-03-25 | 2023-11-07 | Regeneron Pharmaceuticals, Inc. | Citrate buffered VEGF antagonist formulations |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107115294A (en) | 2017-09-01 |
| CN103212075A (en) | 2013-07-24 |
| CN107115294B (en) | 2019-10-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103212075B (en) | A kind of eye drops containing VEGF antagonist | |
| CN110664757B (en) | Nanocrystalline eye drop, preparation method and application thereof | |
| CN109475506A (en) | The composition and method of the eye disease formed with aberrant nascent vessels are treated using Nintedanib | |
| CN102083439A (en) | Methods for using TGF-beta receptor inhibitors or activin-like kinase (ALK) 5 inhibitors A-83-01 and SB-431542 to treat eye disease and wound healing conditions | |
| CN109675038A (en) | Enhance the composition of low concentration atropic category drug safety and clinical efficacy | |
| CN105392494A (en) | Chimeric cytokine formulations for ocular delivery | |
| CN104379129A (en) | Topical ophthalmological pharmaceutical composition containing pazopanib | |
| CN101947309B (en) | Human basic fibroblast growth factor eye drops and preparation method thereof | |
| CN102389392B (en) | Recombinant bovine basic fibroblast growth factor gel for eye use | |
| CN104814924A (en) | Brinzolamide liposome eye preparation and preparation method thereof | |
| CN108295072A (en) | Nintedanib prevents the purposes of eye disease | |
| CN102836171A (en) | Solution for surgery and endoscope washing and preparation method thereof | |
| CN110075148A (en) | Arasaponin extract is preparing the application in eye medicinal preparation | |
| CN103816115B (en) | A kind of pharmaceutical composition containing the fusion protein for suppressing blood vessel hyperplasia and purposes | |
| CN116763727A (en) | Etoposide injection and preparation method thereof | |
| CN102008488B (en) | Triamcinolone acetonide ophthalmic preparation and preparation method thereof | |
| CN104606666B (en) | BFGF bovine basic fibroblast growth factor eye drop | |
| CN112656759B (en) | Rudexilvir eye drops and preparation method and application thereof | |
| CN104721136A (en) | Brinzolamide nanosuspension used for eyes and preparation method thereof | |
| CN107537036A (en) | A kind of pharmaceutical preparation of human vessel endothelium growth factor resisting monoclonal antibody and its application | |
| CN104721145A (en) | Brinzolamide nanoparticle preparation used for eyes and preparation method thereof | |
| Ahmed et al. | Sterile dosage forms loaded nanosystems for parenteral, nasal, pulmonary and ocular administration | |
| Jinapriya et al. | Anaesthetic plus dilating gel improves pupil dilation for cataract surgery | |
| CN105213418A (en) | Preoperative compound eye drops of a kind of ophthalmology and preparation method thereof | |
| CN105012305A (en) | Application of lamivudine and pharmaceutical salt thereof in treating age-related macular degeneration |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |