WO2021129658A1 - 融合蛋白在治疗年龄相关性黄斑变性中的应用 - Google Patents

融合蛋白在治疗年龄相关性黄斑变性中的应用 Download PDF

Info

Publication number
WO2021129658A1
WO2021129658A1 PCT/CN2020/138598 CN2020138598W WO2021129658A1 WO 2021129658 A1 WO2021129658 A1 WO 2021129658A1 CN 2020138598 W CN2020138598 W CN 2020138598W WO 2021129658 A1 WO2021129658 A1 WO 2021129658A1
Authority
WO
WIPO (PCT)
Prior art keywords
eye
administration
ibi302
weeks
individual
Prior art date
Application number
PCT/CN2020/138598
Other languages
English (en)
French (fr)
Inventor
卢淑杰
钱镭
Original Assignee
信达生物制药(苏州)有限公司
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 信达生物制药(苏州)有限公司 filed Critical 信达生物制药(苏州)有限公司
Priority to CA3165887A priority Critical patent/CA3165887A1/en
Priority to CN202080090126.6A priority patent/CN114867489A/zh
Priority to EP20904512.9A priority patent/EP4082559A4/en
Priority to JP2022538928A priority patent/JP2023509880A/ja
Priority to AU2020414152A priority patent/AU2020414152A1/en
Priority to US17/788,181 priority patent/US20230056821A1/en
Publication of WO2021129658A1 publication Critical patent/WO2021129658A1/zh

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • A61K38/1793—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Definitions

  • the present invention relates to the application of a bispecific fusion protein that inhibits the activation of the complement pathway and the vascular endothelial growth factor (VEGF) pathway in the treatment of age-related macular degeneration.
  • the present invention also relates to a pharmaceutical composition containing the fusion protein and a method of using the fusion protein to treat age-related macular degeneration.
  • Age-related macular degeneration is a chronic progressive disease of the central retina. Age-related macular degeneration mostly occurs over the age of 45, and its prevalence increases with age. It is currently an important disease that causes blindness in the elderly. There are about 30 million patients with age-related macular degeneration in the world, and about 500,000 people are blinded by this every year. With the development of my country's economy and the aging of the population, the incidence of AMD in my country is increasing year by year, and it has now become the third leading cause of blindness in my country.
  • AMD is mainly manifested by the decreased ability of retinal pigment epithelial cells (RPE) to phagocytose and digest the outer segmental membrane of the optic cell.
  • RPE retinal pigment epithelial cells
  • the residual disc membranes that have not been completely digested are retained in the basal cell protoplasm and overflow and deposit outside the cell.
  • drusen is formed. Pigment epithelium, Bruch's membrane and visual cells in drusen have varying degrees of degeneration, proliferation or atrophy.
  • the "Age-related Eye Disease Research" group of the National Eye Institute of the United States divides AMD into early, middle and late AMD. Early AMD is mainly characterized by drusen, with high incidence and low blindness.
  • Exudative AMD is also called wet AMD, neovascular AMD, the main pathological feature is increased release of vascular endothelial cell growth factor (VEGF), immature neovascularization between pigment epithelial cells or photoreceptors in the center of the retina, namely choroid Neovascularization (CNV), subretinal rupture and hemorrhage, destruction of retinal pigment epithelium and Bruch membrane, low incidence and high blindness.
  • VEGF vascular endothelial cell growth factor
  • CNV choroid Neovascularization
  • the pathological mechanism of AMD has not been fully elucidated. At present, it is generally accepted that angiogenesis induced by increased VEGF expression is the main cause of wet AMD.
  • the inflammatory response mediated by the mutation of complement genes and abnormal activation of complement is also considered to be an important cause of the onset of AMD.
  • the activation of the complement system has a direct effect on RPE damage. On the one hand, it leads to RPE atrophy and induces the atrophy of central retinal cells, thereby forming GA (atrophic AMD); on the other hand, it destroys the blood-retinal barrier, leading to inflammation and high expression of VEGF. New blood vessels are formed, thereby forming neovascular AMD (exudative AMD).
  • Anti-complement drugs are mainly used for dry AMD with geographic atrophy, but the efficacy of wet AMD with higher blindness is not clear, and it may need to be combined with anti-VEGF drugs to show advantages.
  • the current combination therapy uses anti-complement drugs and anti-VEGF drugs to be administered by intravitreal injection respectively, and there is no anti-complement and anti-VEGF double antibody drugs. Due to the risk of vitreous injection, the patient's compliance is poor when repeated long-term administration. Therefore, this combination therapy increases the risk of injection operation and imposes a huge psychological and economic burden on the patient. Therefore, the development can be carried out at the same time. Drugs that inhibit VEGF and complement have potential clinical value. In addition, complement activation plays an important role in the formation of geographic atrophy. For wet AMD patients with geographic atrophy, the fusion protein of anti-complement and anti-VEGF may have important clinical value.
  • WO2013082563A1 discloses a new fusion protein that inhibits the VEGF pathway and the complement pathway.
  • WO2013082563A1 only verified the activity of the fusion protein in animals, and the research on whether it is effective for human AMD disease is still underway, and considering the complexity of human ocular AMD disease, the operation and administration of ocular drug Due to the limitation and particularity of drug capacity, it is still necessary to conduct in-depth research and improvement on the methods and specific schemes of using such fusion proteins for human AMD disease.
  • the present invention relates to a method of preventing or treating age-related macular degeneration (AMD) in an individual, the method comprising administering to the individual a fusion protein that inhibits the VEGF pathway and the complement pathway.
  • AMD age-related macular degeneration
  • the fusion protein includes the sequence of SEQ ID NO:1:
  • the sequence of the fusion protein is SEQ ID NO: 1.
  • the fusion protein is referred to as IBI302 in this application, and the preparation method is referred to WO2013082563A1.
  • the AMD is wet AMD.
  • the wet AMD has one or more of the following symptoms and signs: decreased vision, distorted vision, central dark spots, dyslexia, swelling of the omentum in the macula, fundus hemorrhage, neovascularization, Scar fibrosis, geographic atrophy, etc., or any combination thereof.
  • the wet AMD has choroid neovascularization (CNV).
  • CNV choroid neovascularization
  • the wet AMD is AMD with the following characteristics:
  • OCT optical coherence tomography
  • the individual is a human individual.
  • the individual is a human individual who has the diseases, symptoms and/or characteristics described in any one of the above embodiments.
  • the individual is a human individual who is a patient with wet AMD and is accompanied by geographic atrophy.
  • the individual has wet AMD with the following characteristics:
  • OCT optical coherence tomography
  • the treatment can achieve one or more of the following effects in the individual:
  • CNV choroidal neovascularization
  • the treatment can achieve one or more of the following effects in the individual:
  • CNV choroidal neovascularization
  • the treatment can achieve one or more of the following effects in the individual:
  • the best corrected visual acuity is improved by at least 1 letter of ETDRS, preferably by at least 5 letters of ETDRS, more preferably by 5 to 35 letters of ETDRS, for example by 5-25 letters of ETDRS, for example 5-10, 5-15, 5-20, 10-15, 10-20, 10-25, 15-20 or 15-25 letters, for example, increase 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 , 34 or 35 letters; and/or
  • the best corrected visual acuity is increased by more than 5%, preferably by more than 10%, for example, by 10%-200%, preferably by 10%-150%, more preferably by 10%-100%, more preferably 20%-100%, more preferably 50%-100%, 60%-100%, 20%-80% or 30%-70%, more preferably 50%-70%, more preferably 60%-70%, such as 5 %, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190% or 200%, and/or
  • the central retinal thickness (CST) is lower than the baseline level by more than 25 ⁇ m, for example, 25-350 ⁇ m, preferably 50-300 ⁇ m, more preferably 50-200 ⁇ m, such as 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 ⁇ m, or more than 5% lower than the baseline level, such as 10%-70%, preferably 10%-60%, more preferably 30%-50%, more preferably 40-50% , Such as 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70%; and/or
  • CNV choroidal neovascularization
  • the area reduced compared to baseline 0.1mm 2 or more e.g. reduced 0.1-20mm 2, preferably 0.2-5mm 2, more preferably 0.3-2mm 2, for example reduce 0.1,0.2,0.3,0.4 , 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 , 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.5, 6, 6.5, 7 , 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5 Or 20mm 2 , or
  • the fusion protein of the present invention is formulated for administration as a liquid pharmaceutical composition.
  • Usable carriers and solvents include water, Ringer's solution, phosphate buffered saline solution, and isotonic sodium chloride solution.
  • sterile, non-volatile oil can also be used as a solvent or suspension medium when appropriate.
  • any mixture of non-volatile mineral oils or non-mineral oils can be used, including synthetic monoglycerides or diglycerides.
  • fatty acids such as oleic acid can also be used in the preparation of injections.
  • the pharmaceutical composition comprising the fusion protein of the present invention is a solution for injection or a dry powder formulation.
  • the composition is a lyophilized powder, which can be reconstituted as an injection in a pharmaceutically acceptable liquid carrier.
  • the pharmaceutically acceptable liquid carrier can be, for example, sterile water, Ringer's solution, phosphate buffered saline solution, isotonic sodium chloride solution and the like.
  • the fusion protein of the invention is administered locally.
  • the fusion protein of the invention is administered by intravenous injection.
  • the fusion protein of the invention is administered intralesional.
  • the pharmaceutical composition comprising the fusion protein of the present invention is directly administered to the eye or eye tissue.
  • the pharmaceutical composition is administered topically to the eye.
  • the pharmaceutical composition is administered by injection into the eye (intraocular injection) or tissue associated with the eye.
  • intraocular injection periocular injection, subretinal injection, intravitreal injection, transdiaphragm injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, subfascial injection, post-eye injection
  • the composition is administered by periocular injection or posterior parascleral delivery.
  • composition can also be administered, for example, to the vitreous body, optic nerve, aqueous humor, sclera, conjunctiva, area between sclera and conjunctiva, retinal choroid tissue, macula, or other areas in or near the eye of an individual.
  • intravitreal injection is used for administration.
  • the frequency of administration of the fusion protein of the present invention is three times a day, twice a day, once a day, once every two days, twice a week, once a week, once every two weeks, once every four weeks, every five weeks. Once a week, once every six weeks, once every seven weeks, once every eight weeks, or once a month, once every two months or longer.
  • the administration cycle of the fusion protein of the present invention can be one week, two weeks, three weeks, one month, two months, three months or longer, and the interval between each administration cycle can be the same or different.
  • the drug is administered once or twice in each dosing cycle, preferably once, and the mode of administration is administration on the first day of each cycle.
  • the single administration dose of the fusion protein of the present invention may be selected from 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye , More preferably 1-5mg/eye, more preferably 2-5mg/eye, such as 0.5-2mg/eye, 1-3mg/eye, 2-4mg/eye, 2-3mg/eye or 3-5mg/eye, for example 0.05 mg/eye, 0.1mg/eye, 0.15mg/eye, 0.2mg/eye, 0.25mg/eye, 0.3mg/eye, 0.35mg/eye, 0.4mg/eye, 0.45mg/eye, 0.5mg/eye, 0.55 mg/eye, 0.6mg/eye, 0.65mg/eye, 0.7mg/eye, 0.75mg/eye, 0.8mg/eye, 0.85mg/eye, 0.9mg/eye, 0.95mg/eye, 1mg/eye, 0.01-10 mg/
  • the dosage mode is preferably directly applied to the eye or ocular tissues of the pharmaceutical composition containing the fusion protein of the present invention, for example, the pharmaceutical composition is locally applied to the eye, or through the eye (intraocular injection) or combined with the eye
  • intraocular injection periocular injection, subretinal injection, intravitreal injection, transdiaphragm injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, ocular muscle
  • the liquid composition is administered by submembrane injection, retro-ocular injection, periocular injection, or posterior parascleral delivery.
  • composition can also be administered, for example, to the vitreous body, optic nerve, aqueous humor, sclera, conjunctiva, area between sclera and conjunctiva, retinal choroid tissue, macula, or other areas in or near the eye of an individual.
  • intravitreal injection is used for administration.
  • the fusion protein of the present invention or the pharmaceutical composition thereof can be administered alone or in combination with other drugs.
  • the individual’s central retinal thickness level Decrease more than 25 ⁇ m, for example, decrease 25-350 ⁇ m, preferably 50-300 ⁇ m, more preferably 50-200 ⁇ m, such as decrease 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 ⁇ m , Or reduced by more than 5% compared to the pre-administration level, for example by 10%-70%, preferably 10%-60%, more preferably 30%-50%, more preferably 40-50%, such as 5%, 10% , 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70%.
  • choroidal neovascularization (CNV) area compared with the pre-administration level of the individual's choroidal neovascularization (CNV) area, at 2 weeks, 3 weeks, or 4 weeks after administration, 5 weeks after administration, after administration 6 weeks, 7 weeks after administration, 8 weeks after administration, 9 weeks after administration, 10 weeks after administration, 3 months after administration, 4 months after administration, or 5 months after administration, the individual
  • the area of choroidal neovascularization (CNV) is reduced by more than 0.1mm 2 , such as 0.1-20mm 2 , preferably 0.2-5mm 2 , more preferably 0.3-2mm 2 , such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7 , 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2 , 3.3,
  • the test compared with the pre-administration level of the individual's best corrected visual acuity (BCVA), at 2 weeks, 3 weeks, or 4 weeks after administration, 5 weeks after administration, 6 weeks after administration, Week, 7 weeks after administration, 8 weeks after administration, 9 weeks after administration, 10 weeks after administration, 3 months after administration, 4 months after administration, or 5 months after administration, the test
  • the best corrected visual acuity (BCVA) of the patient is improved by at least 1 ETDRS letter, preferably at least 5 ETDRS letters, more preferably 5-35 ETDRS letters, for example, 5-25 ETDRS letters, for example 5-10, 5-15, 5-20, 10-15, 10-20, 10-25, 15-20 or 15-25 letters, for example, increase 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 Letter, or increase by more than 5% compared to the level before administration, preferably by more than 10%, for example by 10%-200%, preferably 10%-15
  • the individual after administration, the individual does not experience serious adverse events.
  • the individual after administration, has an adverse event rate comparable to that of the individual receiving a placebo.
  • the individual is a human.
  • the method comprises giving the individual about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye, more Preferably 1-5 mg/eye, more preferably 2-5 mg/eye, such as 0.5-2 mg/eye, 1-3 mg/eye, 2-4 mg/eye, 2-3 mg/eye, or 3-5 mg/eye, such as 0.05 mg/eye Eye, 0.1mg/eye, 0.15mg/eye, 0.2mg/eye, 0.25mg/eye, 0.3mg/eye, 0.35mg/eye, 0.4mg/eye, 0.45mg/eye, 0.5mg/eye, 0.55mg/ Eye, 0.6mg/eye, 0.65mg/eye, 0.7mg/eye, 0.75mg/eye, 0.8mg/eye, 0.85mg/eye, 0.9mg/eye, 0.95mg/eye, 1mg/eye, 1.1mg/eye , 1.2
  • the method comprises giving the individual about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye, more Preferably 1-5 mg/eye, more preferably 2-5 mg/eye, such as 0.5-2 mg/eye, 1-3 mg/eye, 2-4 mg/eye, 2-3 mg/eye, or 3-5 mg/eye, for example, 0.05 mg/eye Eye, 0.1mg/eye, 0.15mg/eye, 0.2mg/eye, 0.25mg/eye, 0.3mg/eye, 0.35mg/eye, 0.4mg/eye, 0.45mg/eye, 0.5mg/eye, 0.55mg/ Eye, 0.6mg/eye, 0.65mg/eye, 0.7mg/eye, 0.75mg/eye, 0.8mg/eye, 0.85mg/eye, 0.9mg/eye, 0.95mg/eye, 1mg/eye, 1.1mg/eye ,
  • the method comprises giving the individual about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye, more Preferably 1-5 mg/eye, more preferably 2-5 mg/eye, such as 0.5-2 mg/eye, 1-3 mg/eye, 2-4 mg/eye, 2-3 mg/eye, or 3-5 mg/eye, such as 0.05 mg/eye Eye, 0.1mg/eye, 0.15mg/eye, 0.2mg/eye, 0.25mg/eye, 0.3mg/eye, 0.35mg/eye, 0.4mg/eye, 0.45mg/eye, 0.5mg/eye, 0.55mg/ Eye, 0.6mg/eye, 0.65mg/eye, 0.7mg/eye, 0.75mg/eye, 0.8mg/eye, 0.85mg/eye, 0.9mg/eye, 0.95mg/eye, 1mg/eye, 1.1mg/eye , 1.2
  • the present invention relates to the use of a fusion protein that inhibits the VEGF pathway and the complement pathway in the preparation of a medicament for the prevention or treatment of age-related macular degeneration (AMD) in an individual.
  • AMD age-related macular degeneration
  • the present invention relates to a fusion protein that inhibits the VEGF pathway and the complement pathway, which is used to prevent or treat age-related macular degeneration (AMD) in an individual.
  • AMD age-related macular degeneration
  • the fusion protein includes the sequence of SEQ ID NO:1:
  • the sequence of the fusion protein is SEQ ID NO: 1.
  • the fusion protein is referred to as IBI302 in this application, and the preparation method is referred to WO2013082563A1.
  • the AMD is wet AMD.
  • the wet AMD has one or more of the following symptoms and signs: decreased vision, distorted vision, central dark spots, dyslexia, swelling of the omentum in the macula, fundus hemorrhage, neovascularization, Scar fibrosis, geographic atrophy, or any combination thereof.
  • the wet AMD has choroidal neovascularization (CNV).
  • CNV choroidal neovascularization
  • the wet AMD is AMD with the following characteristics:
  • OCT optical coherence tomography
  • the individual is a human individual.
  • the individual is a human individual who has the diseases, symptoms and/or characteristics described in any one of the above embodiments.
  • the individual is a human individual who is a patient with wet AMD and is accompanied by geographic atrophy.
  • the individual has wet AMD with the following characteristics:
  • OCT optical coherence tomography
  • the treatment can achieve one or more of the following effects in the individual:
  • CNV choroidal neovascularization
  • the treatment can achieve one or more of the following effects in the individual:
  • CNV choroidal neovascularization
  • the treatment can achieve one or more of the following effects in the individual:
  • the best corrected visual acuity is improved by at least 1 letter of ETDRS, preferably by at least 5 letters of ETDRS, more preferably by 5 to 35 letters of ETDRS, for example by 5-25 letters of ETDRS, for example 5-10, 5-15, 5-20, 10-15, 10-20, 10-25, 15-20 or 15-25 letters, for example, increase 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 , 34 or 35 letters; and/or
  • the best corrected visual acuity is increased by more than 5%, preferably by more than 10%, for example, by 10%-200%, preferably by 10%-150%, more preferably by 10%-100%, more preferably 20%-100%, more preferably 50%-100%, 60%-100%, 20%-80% or 30%-70%, more preferably 50%-70%, more preferably 60%-70%, such as 5 %, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190% or 200%, and/or
  • the central retinal thickness (CST) is lower than the baseline level by more than 25 ⁇ m, for example, 25-350 ⁇ m, preferably 50-300 ⁇ m, more preferably 50-200 ⁇ m, such as 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 ⁇ m, or more than 5% lower than the baseline level, such as 10%-70%, preferably 10%-60%, more preferably 30%-50%, more preferably 40-50% , Such as 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70%; and/or
  • CNV choroidal neovascularization
  • the area reduced compared to baseline 0.1mm 2 or more e.g. reduced 0.1-20mm 2, preferably 0.2-5mm 2, more preferably 0.3-2mm 2, for example reduce 0.1,0.2,0.3,0.4 , 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 , 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.5, 6, 6.5, 7 , 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5 Or 20mm 2 , or
  • the frequency of administration of the fusion protein of the present invention is three times a day, twice a day, once a day, once every two days, twice a week, once a week, once every two weeks, once every four weeks, every five weeks. Once a week, once every six weeks, once every seven weeks, once every eight weeks, or once a month, once every two months or longer.
  • the administration cycle of the fusion protein of the present invention can be one week, two weeks, three weeks, one month, two months, three months or longer, and the interval of each administration cycle can be the same or different.
  • the drug is administered once or twice in each dosing cycle, preferably once, and the mode of administration is administration on the first day of each cycle.
  • the single administration dose of the fusion protein of the present invention is selected from 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye, More preferably 1-5 mg/eye, more preferably 2-5 mg/eye, such as 0.5-2 mg/eye, 1-3 mg/eye, 2-4 mg/eye, 2-3 mg/eye or 3-5 mg/eye, such as 0.05 mg /Eye, 0.1mg/eye, 0.15mg/eye, 0.2mg/eye, 0.25mg/eye, 0.3mg/eye, 0.35mg/eye, 0.4mg/eye, 0.45mg/eye, 0.5mg/eye, 0.55mg /Eye, 0.6mg/eye, 0.65mg/eye, 0.7mg/eye, 0.75mg/eye, 0.8mg/eye, 0.85mg/eye, 0.9mg/eye, 0.95mg/eye, 1mg/eye,
  • the fusion protein of the present invention or the pharmaceutical composition thereof can be administered alone or in combination with other drugs.
  • the individual's central retinal thickness level decreased by more than 25 ⁇ m , such as reducing 25-350 ⁇ m, preferably 50-300 ⁇ m, more preferably 50-200 ⁇ m, such as reducing 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 ⁇ m, or with
  • the level before administration is reduced by more than 5%, for example by 10%-70%, preferably 10%-60%, more preferably 30%-50%, more preferably 40-50%, such as 5%, 10%, 15% , 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70%.
  • the individual compared to the pre-administration level of the individual's choroidal neovascularization (CNV) area, at 2 weeks, 3 weeks, or 4 weeks after administration, 5 weeks after administration, 6 weeks after administration, 7 weeks after administration, 8 weeks after administration, 9 weeks after administration, 10 weeks after administration, 3 months after administration, 4 months after administration, or 5 months after administration, the individual’s choroidal neoplasia angiogenesis (CNV) to reduce the area of 0.1mm 2 or more, e.g.
  • CNV choroidal neoplasia angiogenesis
  • the most Better corrected visual acuity (BCVA) improves at least 1 ETDRS letter, preferably at least 5 ETDRS letter, more preferably 5-35 ETDRS letter, for example 5-25 ETDRS letter, for example 5-10, 5-15 One, 5-20, 10-15, 10-20, 10-25, 15-20 or 15-25 letters, for example, increase 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 letters, or Compared with the level before administration, it is increased by more than 5%, preferably by more than 10%, for example, by 10%-200%, preferably by 10%-150%, more preferably by 10%-10
  • the individual after administration, the individual does not experience serious adverse events.
  • the incidence of adverse events in the individual is comparable to that of the individual receiving a placebo.
  • the individual is a human.
  • the fusion protein is administered to the individual in the following schedule: about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye , More preferably 1-5mg/eye, more preferably 2-5mg/eye, such as 0.5-2mg/eye, 1-3mg/eye, 2-4mg/eye, 2-3mg/eye or 3-5mg/eye single time Dosage, once a day, once every two days, twice a week, once a week, once every two weeks, once every four weeks, once every five weeks or once every six weeks intravitreal or intravenous administration of the fusion protein, and the individual Compared to the pre-administration level of the central retinal thickness level of the individual, the individual’s central retinal thickness level is reduced by more than 25 ⁇ m at six or eight weeks after administration, for example by 25-350 ⁇ m, preferably 50-300 ⁇ m, more preferably 50-200 ⁇ m, For example, reduce 25, 50
  • the fusion protein is administered to the individual in the following schedule: about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye , More preferably 1-5mg/eye, more preferably 2-5mg/eye, such as 0.5-2mg/eye, 1-3mg/eye, 2-4mg/eye, 2-3mg/eye or 3-5mg/eye single time Dosage, once a day, once every two days, twice a week, once a week, once every two weeks, once every four weeks, once every five weeks or once every six weeks intravitreal or intravenous administration of the fusion protein, and the individual Compared with the pre-administration level of the choroidal neovascularization (CNV) area, the individual’s choroidal neovascularization (CNV) area is reduced by more than 0.1mm 2 at six or eight weeks after administration, for example by 0.1-20mm 2 , Preferably 0.2-5mm 2 ,
  • the fusion protein is administered to the individual in the following schedule: about 0.01-10 mg/eye, preferably 0.05-8 mg/eye, more preferably 0.05-6 mg/eye, more preferably 0.5-5 mg/eye , More preferably 1-5mg/eye, more preferably 2-5mg/eye, such as 0.5-2mg/eye, 1-3mg/eye, 2-4mg/eye, 2-3mg/eye or 3-5mg/eye single time Dosage, once a day, once every two days, twice a week, once a week, once every two weeks, once every four weeks, once every five weeks or once every six weeks intravitreal or intravenous administration of the fusion protein, and the individual Compared with the pre-administration level of best corrected visual acuity (BCVA), the individual’s best corrected visual acuity (BCVA) is increased by at least 1 letter of ETDRS, preferably by at least 5 letters of ETDRS, six or eight weeks after administration, More preferably, increase 5-35 ETDRS letters, such as
  • the present invention relates to a single drug dosage unit characterized by comprising a fusion protein that inhibits the VEGF pathway and the complement pathway.
  • the fusion protein includes the sequence of SEQ ID NO:1:
  • the sequence of the fusion protein is SEQ ID NO: 1.
  • the fusion protein is referred to as IBI302 in this application, and the preparation method is referred to WO2013082563A1.
  • the single drug dosage unit contains the following dosage of the fusion protein: 0.01-10 mg, preferably 0.05-8 mg, more preferably 0.05-6 mg, more preferably 0.5-5 mg, more preferably 1-5 mg, more Preferably 2-5mg, such as 0.5-2mg, 1-3mg, 2-4mg, 2-3mg or 3-5mg, for example 0.05mg, 0.1mg, 0.15mg, 0.2mg, 0.25mg, 0.3mg, 0.35mg, 0.4mg , 0.45mg, 0.5mg, 0.55mg, 0.6mg, 0.65mg, 0.7mg, 0.75mg, 0.8mg, 0.85mg, 0.9mg, 0.95mg, 1mg, 1.1mg, 1.2mg, 1.3mg, 1.4mg, 1.5mg , 1.6mg, 1.7mg, 1.8mg, 1.9mg, 2mg, 2.1mg, 2.2mg, 2.3mg
  • the present invention relates to a packaged medicine packaging box, which is characterized by comprising the fusion protein according to any one of the above-mentioned single-dose unit embodiments.
  • the present invention relates to the use of a single drug dosage unit as described above or a kit as described above in the preparation of a medicament for the prevention or treatment of age-related macular degeneration (AMD).
  • AMD age-related macular degeneration
  • the present invention relates to a single drug dosage unit as described above or a kit of kits as described above, which is used for the prevention or treatment of age-related macular degeneration (AMD).
  • AMD age-related macular degeneration
  • the term “comprising” or “including” means including the stated elements, integers or steps, but does not exclude any other elements, integers or steps.
  • the term “comprising” or “including” when used, unless otherwise specified, it also covers the combination of the stated elements, integers or steps.
  • an antibody variable region that "comprises” a specific sequence it is also intended to encompass the antibody variable region composed of the specific sequence.
  • “Individual” includes mammals. Mammals include, but are not limited to, domestic animals (for example, cattle, sheep, cats, dogs, and horses), primates (for example, human and non-human primates such as monkeys), rabbits, and rodents (for example, , Mice and rats). In some embodiments, the individual is a human, including a child, adolescent, or adult.
  • administered in combination refers to the administration of two or more therapeutic agents to treat diseases as described in the present disclosure.
  • Such administration includes co-administration of these therapeutic agents in a substantially simultaneous manner, for example, in a single composition having a fixed ratio of active ingredients.
  • such administration includes co-administration of the respective active ingredients in multiple or separate containers (e.g., tablets, capsules, powders and liquids).
  • the powder and/or liquid can be reconstituted or diluted to the desired dose before administration.
  • such administration also includes the use of each type of therapeutic agent in a sequential manner at approximately the same time or at different times. In either case, the treatment regimen will provide the beneficial effects of the drug combination in the treatment of the conditions or conditions described herein.
  • treatment refers to slowing, interrupting, blocking, alleviating, stopping, reducing, or reversing the progression or severity of an existing symptom, disorder, condition, or disease.
  • prevention includes the inhibition of the occurrence or development of a disease or condition or the symptoms of a particular disease or condition.
  • individuals with a family history are candidates for preventive regimens.
  • prevention refers to the administration of drugs before the onset of signs or symptoms, especially in at-risk individuals.
  • the term "effective amount” refers to the amount or dose of the preparation or fusion protein of the present invention, which produces the desired effect in the treated patient after being administered to the patient in single or multiple doses.
  • the effective amount can be easily determined by the attending physician as a person skilled in the art by considering various factors such as the species of mammal; its size, age and general health; the specific disease involved; the degree or severity of the disease; The response of the individual patient; the specific antibody administered; the mode of administration; the bioavailability characteristics of the administered formulation; the selected dosing regimen; and the use of any concomitant therapy.
  • Therapeutically effective amount refers to the amount that is effective to achieve the desired therapeutic result at the required dose and for the required period of time.
  • the therapeutically effective amount of the preparation, antibody or antibody fragment or conjugate or composition of the present invention can be based on a variety of factors such as disease state, the age, sex and weight of the individual, and the amount of antibody or antibody part that elicits the desired response in the individual. Ability to change.
  • a therapeutically effective amount is also an amount in which any toxic or harmful effects of the formulation, antibody or antibody fragment or conjugate or composition thereof are not as good as the therapeutically beneficial effects.
  • prophylactically effective amount refers to an amount that effectively achieves the desired preventive result at the required dose and for the required period of time. Generally, since the prophylactic dose is used in the subject before or at an earlier stage of the disease, the prophylactically effective amount will be less than the therapeutically effective amount.
  • fusion protein refers to a polypeptide having two or more parts covalently linked together, wherein each of the parts is derived from a different protein.
  • the two or more parts can be connected directly by a single peptide bond or by a peptide linker containing one or more amino acid residues.
  • the two parts and the linker will be in open reading frame with each other and produced using recombinant technology.
  • fusion protein that inhibits the VEGF pathway and the complement pathway includes a complement inhibitory domain (CID), a VEGF inhibitory domain (VID) and a half-life extension domain.
  • CID complement inhibitory domain
  • VID VEGF inhibitory domain
  • a half-life extension domain a complement inhibitory domain
  • the VEGF The fusion protein is selected from SEQ ID NO: 40 in WO2013082563A1, and the "CID", "VID” and “half-life extension domains" refer to WO2013082563A1.
  • central retina located in the center of the retina, is the most sensitive area for vision. Cone cells responsible for photopic vision and color vision are distributed in this area. Therefore, any disease that affects this area will cause a significant decrease in central vision and may also be combined with color vision. Abnormalities, visual distortion, dark spots in the center, etc. The lesions can be manifested as hemorrhage, edema, increase in thickness, and atrophy. In addition, the macula is located at the very center of the central retinal area.
  • choroidal neovascularization refers to the proliferating blood vessels from the choroidal capillaries, which expand through the breach of Bruch's membrane, between Bruch's membrane and the retinal pigment epithelium, or between the neural retina and the retinal pigment epithelium, or located in the retinal pigment epithelium Proliferation and formation between the choroid and many diseases involving the RPE-Bruch membrane-choroid capillary complex can lead to the formation of choroidal neovascularization.
  • best corrected visual acuity refers to the visual acuity measured after the refractive error of the eyeball is completely corrected. Represents the actual visual ability of the eyeball.
  • the term “formulation” or “pharmaceutical composition” refers to a composition comprising at least one active ingredient and at least one inactive ingredient suitable for administration to animals, preferably mammals (including humans).
  • "Liquid formulation” or “liquid composition” refers to a formulation in liquid form.
  • the liquid composition of the present invention comprises (i) the fusion protein of the present invention; (ii) a buffer; and (iii) a vehicle.
  • the composition of the formulation of the present invention may be as shown in the above embodiments related to liquid pharmaceutical compositions.
  • the liquid preparation of the present invention is preferably an injection.
  • “Pharmaceutically acceptable carrier” refers to ingredients other than the active ingredient in a pharmaceutical preparation, which are not toxic to the subject.
  • Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers or preservatives.
  • buffering agent refers to a pH buffering agent.
  • the buffer is selected from histidine, glutamate, phosphate, acetate, citrate, and tris.
  • the term "vehicle” refers to a liquid used to dissolve or suspend active ingredients and inactive ingredients to form a liquid formulation.
  • the solvents that can be used in the present invention include, but are not limited to, water for injection, and organic solvents for injection include, but are not limited to, injection oil, ethanol, propylene glycol, etc., or a combination thereof.
  • single drug dosage unit refers to a single drug dosage form containing the fusion protein of the present invention to be administered to a patient at the time of administration, such as vials, ampoules, prefilled needles or prefilled forms for injection Syringe containing a solution or lyophilized powder of the drug.
  • Figure 1 depicts the inhibitory effect of IBI302 on VEGF-A-induced HUVEC proliferation.
  • Figure 2 depicts the inhibitory effect of IBI302 on hemolysis mediated by the classical pathway of complement.
  • Figure 3 depicts the inhibitory effect of IBI302 on the alternative pathway of complement mediated hemolysis.
  • Figure 4 depicts the inhibitory effect of each dose of IBI302 on CNV. *P ⁇ 0.05vs PBS group; #P ⁇ 0.05vs IBI302 1, 3 or 5 ⁇ g group.
  • FIG. 5 illustrates that IBI302 inhibits CNV formation and leakage. *P ⁇ 0.05vs PBS group; #P ⁇ 0.05vs IBI302 10 ⁇ g group.
  • Figure 6 depicts the effect of IBI302 on inhibiting inflammatory cell aggregation. *P ⁇ 0.05vs PBS group; #P ⁇ 0.05vs IBI302 10 ⁇ g group.
  • Figure 7 depicts the effect of IBI302 on the fundus fluorescence leakage of the rhesus monkey CNV model (fluorescence contrast).
  • Figure 8 depicts a representative image of choroidal CD31 immunohistochemical staining.
  • Figure 9 depicts a representative image of choroidal C5b-9 immunohistochemical staining.
  • Figure 10 depicts the drug-time curve of serum concentration in rhesus monkeys after intravenous injection and intravitreal injection of IBI302.
  • Figure 11 depicts the concentration-drug-time curve in aqueous and vitreous fluid of rhesus monkeys after intravitreal injection of IBI302.
  • Figure 12 depicts the drug-time curve of the lens, cornea, iris, retina, choroid, optic nerve, and sclera after intravitreal injection of IBI302 in rhesus monkeys.
  • Figure 13 depicts the average blood concentration-time curve of rhesus monkeys after the first intravenous injection and intravitreal injection of different doses of IBI302.
  • Figure 14 depicts the average blood concentration-time curve of rhesus monkeys after the last intravenous injection and intravitreal injection of different doses of IBI302.
  • FIG. 15 depicts a schematic diagram of the dose escalation of the phase I clinical study described in Example 5.
  • FIG. 15 depicts a schematic diagram of the dose escalation of the phase I clinical study described in Example 5.
  • FIG. 16 depicts a schematic diagram of the research process of the phase I clinical study described in Example 5.
  • Figure 17 depicts the change trend of the number of correctly identified letters shown in the initial trial results of the phase I clinical study described in Example 5 and the comparison with the baseline.
  • Figure 18 depicts the central retinal thickness (CST) change trend and the baseline comparison ( ⁇ CST) shown in the initial trial results of the phase I clinical study described in Example 5.
  • Figure 19 depicts the changes over time in the BCVA (number of letters of EDTRS) in each dose group in the overall study results of the phase I clinical study described in Example 5, where the data before the first administration of the study drug (including the data during the screening period) is used The last non-missing value of is used as the baseline.
  • Figure 20 depicts the change over time of the BCVA (number of letters of EDTRS) in each dose group in the overall study results of the phase I clinical study described in Example 5 compared with baseline, where the change in value before the first administration of the study drug (including The last non-missing value of the data in the screening period was used as the baseline.
  • the change value is defined as the average of the difference between the measured value of each visit and the baseline.
  • Figure 21 depicts the overall study results of the phase I clinical study described in Example 5 in the ophthalmological OCT examination of the central area of the macular thickness over time in each dose group, wherein the first administration of the study drug (including the data during the screening period) The last non-missing value is used as the baseline.
  • Figure 22 depicts the changes over time in the overall study results of the phase I clinical study of each dose group in the ophthalmic OCT examination of the central area of the macula compared to the baseline, in which the value before the first administration of the study drug ( Including the data of the screening period) the last non-missing value as the baseline.
  • the change value is defined as the average of the difference between the measured value of each visit and the baseline.
  • Biacore T200 was used to investigate the affinity of IBI302 with related ligands, and the affinity of IBI302 with VEGF family, C3b and C4b, as shown in Table 1.
  • the affinity of IBI302 and VEGF family cytokines is comparable to that of aflibercept injection, and is significantly stronger than that of bevacizumab injection.
  • the affinity of IBI302 and C4b is comparable to CR1; the affinity of IBI302 and C3b is slightly weaker than CR1, possibly because the CID end of IBI302 is only part of the domain of CR1, but it still reaches the nM level.
  • IBI302 inhibits the proliferation of HUVEC induced by VEGF-A
  • HUVEC cells show different proliferation ability according to the amount of free VEGF-A165.
  • the number of HUVEC viable cells is determined by the CCK-8 color reagent kit. In order to determine the inhibitory efficiency of IBI302 on HUVEC cell proliferation. At the same time, aflibercept and bevacizumab injection were used as the reference substance.
  • IBI302 can significantly inhibit the proliferation of HUVEC stimulated by VEGF, similar to the activity of aflibercept, and slightly stronger than bevacizumab injection. See Table 2 and Figure 1.
  • IBI302 can inhibit the hemolysis of human serum complement-induced sensitized sheep red blood cells in a concentration-dependent range from 0.02 to 1080.26 nM, with a median effective concentration (EC50) value of 2.514 nM.
  • the EC50 value of CR1 is 2.254 nM. It suggests that IBI302 can significantly inhibit the classical pathway of complement, similar to CR1. The specific results are shown in Figure 2.
  • IBI302 can inhibit the hemolysis of rabbit red blood cells induced by human serum complement in a concentration-dependent range from 1.50 to 383.39 nM, with an EC50 value of 14.59 nM.
  • the EC50 value of CR1 is 15.30 nM. It is suggested that IBI302 can significantly inhibit the alternative pathway of complement, similar to CR1. The specific results are shown in Figure 3.
  • the experiment uses hRPE cells, which are divided into normal group (nor), model group (ctr), aflibercept group (VEGF-Trap), CR1 group, and IBI302 group. All groups except the normal group were added t-BHP and 10% normal human serum, and the normal group was added an equal volume of PBS. The drug treatment group was added with the corresponding drugs, and the final concentration of each drug was 1 ⁇ g/ml, and the normal group and the model group were added with equal volumes of PBS. After culturing for 4 hours, the barrier function of hRPE monolayer cells was tested.
  • VEGF vascular endothelial growth factor
  • chemokine ligand 2 Cyhemokine (C-C motif) ligand 2, CCL2
  • C3a C5a
  • MAC secreted by hRPE cells were detected by ELISA method.
  • the transepithelial resistance (TER) values of oxidative damage hRPE in the model group (ctr), aflibercept group, CR1 group, and IBI302 group were 54.01%, 64.64%, 63.48%, and 63.48% of the normal group (nor) hRPE, respectively. 75.90%.
  • the TER of the aflibercept group, the CR1 group and the IBI302 group were significantly improved, and the IBI302 group was also significantly different from the aflibercept group and the CR1 group, as shown in Table 3.
  • the concentration of hRPE secreted VEGF was significantly reduced in the aflibercept group, CR1 group and IBI302 group.
  • the VEGF concentration in the IBI302 group was significantly lower.
  • the CCL2 secreted by hRPE in the CR1 group and IBI302 group was significantly lower than that in the model group; while the CLL2 secreted by hRPE in the aflibercept group was not statistically different compared with the model group.
  • the concentration of CCL2 in the IBI302 group was significantly lower, as shown in Table 4 and Table 5.
  • IBI302 has a protective effect on the barrier function of hRPE cells from oxidative damage caused by complement activation.
  • the mechanism may be by simultaneously inhibiting the expression of VEGF and complement activation components C3a, C5a and MAC, further reducing the CCL2 secreted by hRPE. And VEGF.
  • mice were used for laser photocoagulation at 4 points.
  • the mice included in the study were randomly divided into 6 groups, 2 (4 eyes) mice in each group: laser-made modules, phosphate buffer ( PBS) treatment group, IBI302 1 ⁇ g/ ⁇ l, 3 ⁇ g/ ⁇ l, 5 ⁇ g/ ⁇ l, 10 ⁇ g/ ⁇ l dose group.
  • PBS phosphate buffer
  • IBI302 1 ⁇ g/ ⁇ l, 3 ⁇ g/ ⁇ l, 5 ⁇ g/ ⁇ l, 10 ⁇ g/ ⁇ l dose group.
  • 1 ⁇ l of each dose of IBI302 or PBS were injected into the vitreous cavities of each group of choroidal neovascularization (CNV) mice.
  • CNV choroidal neovascularization
  • fluorescein isothiocyanate-dextran (FITC-Dextran) orbital sinus perfusion and choroidal spreading were performed to observe the effect of each dose of IBI302 on the area of CNV induced by laser in mice.
  • mice Further experimental groups are: IBI302 group, aflibercept (VEGF-Trap) group, CR1 group, PBS group. Both IBI302 and the control drug were 10 ⁇ g/ ⁇ l. Immediately after laser photocoagulation, 1 ⁇ l of each group's samples were injected into the vitreous cavity of each group of CNV mice. On the 7th day after laser photocoagulation, the mice underwent fundus angiography to observe the condition of CNV. The mice were anesthetized with chloral hydrate and then dilated with compound tropine.
  • ELISA was used to detect the concentration of VEGF, CCL2, TNF- ⁇ , C3a and C5a protein in RPE-choroid.
  • IBI302 The inhibitory effect of different doses of IBI302 on CNV: On the 7th day after laser photocoagulation, the mouse orbital venous sinus was perfused with FITC-Dextran and choroidal pavement showed that the neovascularization showed petal-like structure. There was no statistical difference in the CNV area between the laser module (laser group) and the PBS group. Compared with the PBS group, IBI302 1 ⁇ g group, 3 ⁇ g group, 5 ⁇ g group, 10 ⁇ g group CNV area were reduced by 14.27%, 48.79%, 62.46%, 68.83% compared with PBS group. IBI302 The difference in CNV area between the 10 ⁇ g group, the 1 ⁇ g group, the 3 ⁇ g group, and the 5 ⁇ g group was statistically significant, as shown in Table 9 and Figure 4.
  • 10 ⁇ g/eye of IBI302 Compared with the PBS control group, 10 ⁇ g/eye of IBI302, aflibercept and CR1 all inhibited the area and leakage of CNV. Compared with aflibercept and CR1 group, 10 ⁇ g/eye IBI302 can significantly inhibit CNV area and leakage, see Table 10, 11 and Figure 5.
  • the inhibitory effect of IBI302 on laser-induced VEGF production in CNV mice On the 3rd day after laser photocoagulation, 10 ⁇ g/eye IBI302, aflibercept and CR1 can inhibit laser-induced VEGF in CNV mice compared with PBS group. generate. Compared with the aflibercept group and the CR1 group, the difference in the effect of IBI302 on inhibiting VEGF production was statistically significant, as shown in Table 12.
  • IBI302 inhibits laser-induced complement activation in CNV mice: On the 3rd day after laser photocoagulation, choroidal immunofluorescence spreading showed that compared with the PBS group, aflibercept group, and CR1 group, 10 ⁇ g/eye IBI302 can significantly inhibit MAC in the choroid Of deposition. Twelve hours after laser photocoagulation, the expression levels of C3a and C5a in the RPE-choroid complex were detected by ELISA and found that, compared with the model group, the expression of C3a and C5a in the CR1 group and IBI302 group was significantly reduced.
  • IBI302 Inhibition of IBI302 on the inflammatory factors CCL2 and TNF- ⁇ in laser-induced CNV mice: ELISA was used to detect the inflammatory factors CCL2 and TNF- ⁇ in the RPE-choroid complex at 12h and 3 days after laser photocoagulation, respectively The expression level. Compared with the PBS group, the expressions of inflammatory factors CCL2 and TNF- ⁇ in the 10 ⁇ g/eye CR1 group and IBI302 group were significantly reduced. The concentrations of CCL2 and TNF- ⁇ in the IBI302 group were significantly lower than those in the aflibercept group, as shown in Table 15 and Table 16.
  • IBI302 The inhibitory effect of IBI302 on laser-induced inflammatory cell aggregation in CNV mice: Choroidal pavement immunofluorescence was used to detect the aggregation of neutrophils and macrophages in the choroid on the 1st and 3rd day after laser photocoagulation. IBI302 can significantly inhibit the deposition of neutrophils and macrophages in the choroid, compared with the PBS group and aflibercept group, there are statistical differences, see Table 17, Table 18 and Figure 6.
  • IBI302 can inhibit the formation and leakage of laser-induced CNV in mice.
  • the possible mechanism is that by simultaneously inhibiting the expression of VEGF and the active ingredients C3a, C5a, MAC and CCL2, TNF- ⁇ after complement activation, inhibiting macrophages and Infiltration of neutrophils, thereby inhibiting VEGF secretion and angiogenesis.
  • IBI302 inhibits laser-induced choroidal neovascularization in rhesus monkeys
  • the experiment uses laser photocoagulation around the fovea of the macula of the fundus of rhesus monkeys to induce choroidal neovascularization in the fundus, establishing an animal model similar to human choroidal neovascularization.
  • Selected 25 monkeys (both male and female) with successful binocular modeling were divided into 5 groups, namely the model control group, bevacizumab injection 1.25mg/eye group, IBI302 0.25mg/eye group, IBI302 0.5mg/eye group , IBI302 1.25mg/eye group, 5 monkeys in each group, both male and female. See Table 19 for details. 21 days after photocoagulation, the monkeys in each group were injected with corresponding drugs at a rate of 50 ⁇ l/eye through the vitreous body.
  • Fluorescence contrast and OCT examination There was no obvious change in the degree of fluorescein leakage in the monkeys of the model control group at 14 and 28 days after the administration, and the area of the fluorescent spots was not significantly reduced. The fluorescein leakage in some monkey eyes even increased over time. the trend of. There was no significant change in the number of grade 4 fluorescent spots compared with before administration. 11 and 25 days after the administration, the thickness of the retina was not significantly reduced by OCT, the local Bruch’s membrane structure was not repaired, the choroidal neovascularization was still there, and highly reflective light clusters were visible.
  • the leakage area of the bevacizumab injection group 14 and 28 days after administration was significantly smaller than that before administration, and there was a significant difference compared with the model control group (P ⁇ 0.05).
  • the number of grade 4 fluorescent spots was compared with that before administration. The ratio is significantly reduced.
  • the retinal thickness at the highest point of all eyeball lesions was significantly reduced by OCT, and even approached or reached the pre-modeling level. Compared with the control group, the difference was statistically significant (P ⁇ 0.05).
  • IBI302 0.25mg/eye group, 0.5mg/eye group and 1.25mg/eye group monkeys 14 and 28 days after administration, the leakage area was significantly smaller than before administration, and there was a significant difference compared with the model control group (P ⁇ 0.05) And the improvement of the leakage area at 28 days after administration was also significantly higher than that of the positive control group (P ⁇ 0.05).
  • the number of grade 4 fluorescent spots was significantly reduced compared to before administration.
  • the thickness of the retina in OCT examination was significantly reduced at 11 and 25 days after administration, and even approached or reached the level before modeling. Compared with the model control group, the difference was statistically significant (P ⁇ 0.05). Compared with the positive control group, each The index difference was not statistically significant (P>0.05).
  • VEGF detection 14 and 28 days after administration, the serum VEGF concentration of 1 monkey in the Bevacizumab injection 1.25 mg/eye group and the model control group was measured at the value near the lower limit of quantification, and the other time points were low At the lower limit of quantification. 29 days after administration, the concentration of VEGF in the aqueous humor of the monkeys in the bevacizumab injection 1.25 mg/eye group was lower than that of the control group, and the difference was statistically significant (P ⁇ 0.05). The VEGF concentrations in the aqueous humor of the monkeys in each group of IBI302 were all Below the lower limit of quantification. The specific results are shown in Table 23.
  • HE staining results showed that the model control group had mild to moderate focal retinal/choroidal fibrous tissue hyperplasia and mild to mild focal neuroepithelial detachment.
  • the severity of the above-mentioned lesions in each group of IBI302 and the bevacizumab injection 1.25 mg/eye group was less than that of the model control group.
  • IBI302 can significantly reduce the leakage of fluorescein, the area of fluorescent spots and the number of fluorescent spots at the photocoagulation site, reduce the thickness of the retina at the lesions of fluorescent spots, and restore the pigment epithelium Complete and continuous, reduce the level of VEGF in aqueous humor, reduce the proliferation of fibrous tissue, and inhibit the expression of CD31 and C5b-9 in the choroid.
  • IBI302 has a better effect than clinical dose bevacizumab.
  • mice 50 rhesus monkeys were randomly assigned to 5 groups, the control group, IBI302 intravenous injection 8mg/monkey group, IBI302 vitreous injection 0.5mg/eye group, IBI302 vitreous injection 2mg/eye group and IBI302 vitreous injection 4mg/eye group. There are 10 animals in each group, half male and half male. During the administration period and the recovery period of drug withdrawal, observe the general behavior, mental state, respiratory state, etc.
  • the experimental results showed that during the administration period and during the recovery period of drug withdrawal, the rhesus monkeys in each group were generally in good condition, with normal autonomous activities, and no behavioral abnormalities related to the test article were observed; during the study period, the rhesus monkeys in each group had no abnormalities in body temperature . 6, 24, 48, 120 hours after the first administration, about 6 hours after the last administration, the recovery period is 4 weeks and the recovery period is over, IBI302 monkey heart rate, QT and corrected QT interval, PR interval, RR interval , P wave time, QRS wave time and other electrocardiogram indicators, systolic blood pressure, diastolic blood pressure, mean blood pressure, and visual observation of the respiratory status showed no drug-related abnormal changes.
  • IBI302 0.5mg/eye, 2mg/eye or 4mg/eye in rhesus monkeys or intravenous IBI302 8mg/monkey has no significant effect on the central nervous system, respiratory system and cardiovascular system.
  • IBI302 can specifically bind to the VEGF family through the VID end to inhibit the proliferation of HUVEC induced by VEGF-A; on the other hand, it can specifically bind to C3b and C4b through the CID end to inhibit hemolysis mediated by the alternative complement pathway and the classical pathway. It shows that IBI302 is a dual-target specific monoclonal antibody with a clear target and a clear mechanism of action.
  • IBI302 double-blocks the signal pathway mediated by VEGF-A and complement, inhibits t-BHP from stimulating hRPE cells to release VEGF-A, CCL2, C3a, C5a and MAC to play a protective effect on hRPE cells; inhibits laser-induced choroidal angiogenesis in mice Express VEGF, CCL2, TNF- ⁇ , C3a, C5a and MAC, reduce choroidal angiogenesis; inhibit laser-induced choroidal angiogenesis Rhesus monkeys express VEGF, CD31 and C5-b9, reduce choroidal angiogenesis and neovascular leakage, and reduce the retina The thickness protects the pigmented epithelium, indicating that IBI302 is effective.
  • Dual targets IBI302 specifically binds VEGF-A165, VEGF-A121 and PIGF through the VID end on the one hand, and specifically binds C3b and C4b through the CID end. It is a dual-targeted drug with a clear target.
  • the CID of IBI302 can inhibit the activation of the classical and alternative pathways of complement by specifically binding to C3b and C4b, and reduce the inflammatory response mediated by complement activation; the VID end can bind to the VEGF family to block VEGF-mediated
  • the guided signal pathway inhibits the survival and proliferation of vascular endothelial cells, thereby inhibiting angiogenesis, reducing vascular permeability, and reducing vascular leakage.
  • IBI302 has a significant protective effect on hRPE; in laser-induced mice and rhesus monkey choroidal angiogenesis models, IBI302 can produce significant anti-angiogenesis and infiltration The leakage effect suggests that IBI302 has a very good clinical effect.
  • IBI302 is better than aflibercept or bevacizol in t-BHP-induced hRPE oxidative stress models, laser-induced mice, and rhesus monkey choroidal angiogenesis models. The anti-efficacy is better, suggesting that IBI302 may be more effective than the commercially available anti-VEGF drugs alone.
  • IBI302 Combining the preclinical pharmacological activity of IBI302 to date and the clinical application of similar drugs, IBI302 is clinically intended for the treatment of wet age-related macular degeneration (AMD).
  • AMD wet age-related macular degeneration
  • aqueous humor, vitreous fluid, lens, cornea, iris, retina, choroid, sclera, optic nerve and serum were taken for 4h, 24h, 72h, 168h, 336h, 504h after administration.
  • the detailed experimental design is shown in Table 26.
  • the half-life T1/2 of the drug was significantly longer than that of intravenous injection.
  • the eye drug concentration is significantly higher than the serum drug concentration, and the risk of systemic toxicity is small.
  • the drug is mainly distributed in the vitreous, retina and choroid of the ocular tissue, and regardless of the terminal elimination half-life T1/2 or the average residence time MRT, the drug is both in the retina and choroid. It can stay for a longer period of time and thus play a role.
  • IBI302 In the 4-week toxicity study of repeated administration of IBI302 in rhesus monkeys, 50 rhesus monkeys were randomly assigned to 5 groups, the control group, IBI302 8mg/intravenous injection group, IBI302 0.5mg/eye vitreous injection group, IBI302 2mg/ The intraocular vitreous injection group and the IBI302 4mg/ ocular vitreous injection group were administered once every 2 weeks for three consecutive administrations. Each group had 10 animals, half male and half male. In the control group, blood was collected before the first and last administration, and 24 hours after the administration, and serum was prepared.
  • IBI302 8mg/vessel intravenous injection group blood was collected before the first and last administration, 5min, 1h, 4h, 8h, 24h, 48h, 72h, 120h, 168h, and 336h after the administration, and serum was prepared.
  • IBI302 intravitreal injection administration group blood was collected before the first and last administration, and at 1h, 4h, 8h, 24h, 48h, 72h, 120h and 168h after the administration, and serum was prepared.
  • the established ELISA method was used to detect the concentration of IBI302 in serum.
  • the specific experimental design is shown in Table 29.
  • IBI302 is an antibody drug and does not require metabolic research. Therefore, IBI302 metabolism-related research has not been carried out.
  • the metabolites of IBI302 are expected to be peptides or amino acids.
  • the metabolic rate of IBI302 in the local (eye) area is significantly lower than that of the whole body, and the local drug concentration in the eye is significantly higher than the serum drug concentration.
  • Most drugs are restricted to the eye, and the risk of systemic toxicity is relatively high. small.
  • the ocular exposure of the drug indicates that the drug is most distributed in the vitreous humor, retina, and choroid, thus exerting a drug effect.
  • the drug exposure and peak concentration increase with the increase of the dose, showing a certain dose correlation.
  • the drug exposure in the serum after the last injection has a different degree than the exposure after the first dose.
  • the reduction in immunogenicity showed that some animals produced anti-antibodies, and it also showed that there was no accumulation after repeated administration.
  • Tissue distribution After intravitreal injection of IBI302, most of the drugs are restricted to the eye, and the local drug concentration in the eye is significantly higher than the serum drug concentration, suggesting that IBI302 mainly exerts pharmacological activity in the eye and has a low risk of systemic toxicity. At the same time, the distribution of drugs in the vitreous humor, retina and choroid is the most, which is in line with the design goal of drugs mainly acting on the retina and choroid.
  • Rhesus monkeys were given IBI302 at a dose of 2mg/eye or 4mg/eye through a single intravitreal injection. After 14 days of observation, no obvious toxic reaction was observed in all monkeys, indicating that rhesus monkeys were given a single dose of 2mg/eye or 4mg/eye through vitreous injection The IBI302 can be well tolerated.
  • mice 50 rhesus monkeys were randomly assigned to 5 groups, the control group, IBI302 intravenous injection 8mg/monkey group, IBI302 vitreous injection 0.5mg/eye group, IBI302 vitreous injection 2mg/eye group and IBI302 vitreous injection 4mg/eye group. There are 10 animals in each group, half male and half male.
  • the control group and the IBI302 4mg/eye group were given 0.9% sodium chloride injection and 40mg/ml IBI302 by intravitreal injection of 100 ⁇ l/eye respectively, and 50 ⁇ l/eye for the IBI302 0.5mg/eye group and 2mg/eye group.
  • the volume of 10mg/ml and 40mg/ml of IBI302 were given by intravitreal injection in both eyes.
  • the drug is administered once every 2 weeks for 3 times in a row, and the recovery period of drug withdrawal is 74 days.
  • the specific experimental design is shown in Table 32.
  • Intravenous group and each vitreous injection group Rhesus monkey weight, body temperature, food intake, hematology, blood biochemistry, urine index, bone marrow index, ECG blood pressure, organ weight and organ coefficient, and all organs except eyeball and optic nerve Histopathological examination of organs showed no abnormal changes.
  • Intravitreal injection of each dose of IBI302 can cause different degrees of intraocular inflammation in rhesus monkeys, mainly as follows:
  • 2mg/eye group during the administration period, 1/10 monkeys (starting 3 days after the second administration of the left eye; 3 days after the third administration of the right eye) indirect ophthalmoscope examination revealed anterior chamber exudation and glass weight Opacity, invisible eye fundus or white sheath-like changes around retinal veins and perivascular hemorrhage; Slit lamp examination revealed anterior chamber effusion, pupil dilation, bulbar conjunctival edema, mixed congestion, corneal opacity, aqueous humor flash, iris texture Unclearness, edema, or unclear iris texture, edema; intraocular pressure was significantly reduced; WBC, MONO, NEU, and NEU% were significantly increased; gross anatomy revealed gray-white changes in the pupil areas of the anterior chamber of the eyeball.
  • the 2/4 monkeys In the 4th week of the recovery period, the 2/4 monkeys have white sheath-like changes around the branch vessels of the retina, and basically recovered by the 6th week of the recovery period; the female monkeys at the 4th and 7th week of the recovery period have reduced the amplitude of the 3.0 (Cone-R) b wave; pathological examination only Individual eyes have slight infiltration of optic nerve, ciliary body or retinal monocytes.
  • 4mg/eye group During the administration period, 1/10 monkeys (both eyes, 6 days after the third administration) found opacity of the vitreous in both eyes with indirect ophthalmoscope; Slit lamp examination revealed that the anterior chamber was exuded and the vitreous was opaque. Another monkey, one monkey, had anterior chamber exudation in his left eye one day after the first administration. At the end of the dosing, the female monkey's light adaptation 3.0 (Cone-R) b wave and dark adaptation 3.0 (Max-R) a wave amplitude decreased.
  • Pathological examination revealed eye mononuclear cell infiltration, vitreous degeneration, retinal edema, and slight optic nerve mononuclear cell infiltration.
  • Rhesus monkeys were given IBI302 at a dose of 0.5 mg/eye, 2 mg/eye or 4 mg/eye or 8 mg/eye intravenously via vitreous injection once every two weeks for three consecutive times, and recovered for 74 days. No systemic toxicity was seen at each dose, only some intraocular inflammation was found in individual animals. The recovery period was over, the intraocular inflammation in each group was basically restored by ophthalmological examination, and the pathological changes of the ocular tissues were obviously restored.
  • IBI302 is an antibody drug, and its metabolites are peptides or amino acids, which will not affect DNA and other genetics. Because the substance has an impact, there is no need to carry out genotoxicity research, so IBI302 genotoxicity related research has not been carried out.
  • This product is administered by intravitreal injection, the system exposure is small, and it is aimed at people over 50 years of age with indications. Therefore, no carcinogenicity research has been carried out.
  • This product is administered by intravitreal injection, and the system exposure is small. For the indication population over 50 years old, there is little concern about the safety risk of reproductive toxicity, so no reproductive toxicity research has been carried out.
  • Japanese big-eared white rabbits were given 0.5 mg/eye or 2 mg/eye (concentrations of 10 mg/ml or 40 mg/ml, respectively) once a week for 4 consecutive weeks with IBI302, which was not irritating to rabbit eyes.
  • 50 rhesus monkeys were randomly assigned to 5 groups, the control group, IBI302 intravenous injection 8mg/monkey group, IBI302 vitreous injection 0.5mg/eye group, IBI302 vitreous injection 2mg/eye group and IBI302 vitreous injection 4mg/eye group. There are 10 animals in each group, half male and half male. The drug is administered once every 2 weeks for 3 times in a row, and the recovery period of drug withdrawal is 74 days.
  • the immunogenicity study results showed that after the last administration, the circulating immune complex (CIC) of the female and male monkeys in the vitreous injection 2 mg/eye and 4 mg/eye group increased. In the 4th week of the recovery period, the vitreous injection 4 mg/eye group female and male Monkey CIC also shows an upward trend. In each dose group, some animals produced anti-drug antibodies. The results of biological activity analysis showed that some anti-drug antibodies may be neutralizing antibodies, indicating that IBI302 has a certain degree of immunogenicity.
  • CIC circulating immune complex
  • 50 rhesus monkeys were randomly assigned to 5 groups, the control group, IBI302 intravenous injection 8mg/monkey group, IBI302 vitreous injection 0.5mg/eye group, IBI302 vitreous injection 2mg/eye group and IBI302 vitreous injection 4mg/eye group. There are 10 animals in each group, half male and half male. The drug is administered once every 2 weeks for 3 times in a row, and the recovery period of drug withdrawal is 74 days.
  • the conventional immunohistochemical method was used to investigate the immune cross-reaction of IBI302 with normal human and cynomolgus monkey tissues.
  • the experimental results showed that: (1) Biotinylated IBI302 (Biotin-IBI302) and biotinylated aflibercept (Biotin-ZALTRAP) were used in recombinant human VEGF-A165 protein smears under the conditions of 2 ⁇ g/ml and 20 ⁇ g/ml concentration.
  • Rhesus monkeys were given a single intravitreal injection of IBI302 at a dose of 2 mg/eye or 4 mg/eye. No obvious toxicity was seen, and the tolerability was good. Rhesus monkeys were given IBI302 at a dose of 0.5 mg/eye, 2 mg/eye, or 4 mg/eye or 8 mg/eye by intravenous injection once every 2 weeks for 3 consecutive times, and recovered for 74 days. The results showed that there was no obvious systemic toxicity. Intravitreal injection of IBI302 at each dose can cause different degrees of intraocular inflammation in monkeys, and corresponding inflammatory pathological changes of the eye and optic nerve can be seen.
  • IBI302 Intraocular inflammation caused by intravitreal injection of IBI302 is related to the production of anti-drug antibodies and the formation of immune complexes. Considering that IBI302 is a fully human monoclonal antibody, anti-drug antibodies and immune complexes may appear clinically. It is lower, so the risk of endophthalmitis is lower, but it is still reminded to pay close attention in clinical research.
  • IBI302 has no obvious cross-reaction with normal human tissues and cynomolgus monkey tissues, suggesting that IBI302 has no obvious interaction with normal human tissues, and the target-related toxicity is very low.
  • Example 5 Phase I clinical study of the tolerability and safety of a single dose in patients with wet age-related macular degeneration (AMD)
  • the doses of 0.5mg/eye, 2mg/eye and 4mg/eye can be drawn directly from the existing specifications of the study drug.
  • the volume of medicine to be drawn and the corresponding solvent volume for each dose group are as follows:
  • Dose group Drug specifications Drug volume drawn Dissolved media volume Extract volume (intraocular injection) 0.05mg/eye 0.2ml: 2mg 90 ⁇ l 760 ⁇ l 50 ⁇ l 0.15mg/eye 0.2ml: 8mg 60 ⁇ l 760 ⁇ l 50 ⁇ l 0.5mg/eye 0.2ml: 2mg 50 ⁇ l - 50 ⁇ l 1mg/eye 0.2ml: 8mg 760 ⁇ l 760 ⁇ l 50 ⁇ l 2mg/eye 0.2ml: 8mg 50 ⁇ l - 50 ⁇ l 4mg/eye 0.2ml: 8mg 100 ⁇ l - 100 ⁇ l
  • This study is a multi-center, open, dose-escalation study design.
  • Patients with active subfoveal choroidal neovascularization (CNV) secondary to wet AMD can be included in this clinical study.
  • CNV subfoveal choroidal neovascularization
  • Each subject should not use both eyes as study eyes.
  • Eligible subjects received intravitreal injection of study drug on the first day of the study (D1). After the administration, the subject should receive safety monitoring in the research center until D2, and confirm that there is no clinically significant abnormality before the subject can leave the research center. On D3-D6, D8, D15, D29, D43, the subject should return to the hospital for follow-up. See Figure 15 for a schematic diagram of dose escalation.
  • Dose-limiting toxicity is defined as any of the following adverse events (AE) related to IBI302 that occurred during the DLT observation period:
  • the intraocular pressure continues to increase ⁇ 10mmHg for more than 60min;
  • Intracranial hemorrhage or other clinically significant bleeding in other parts of the eye are intracranial hemorrhage or other clinically significant bleeding in other parts of the eye;
  • SAE serious adverse event
  • Appendix 2 Criteria for the classification of expected ocular adverse events
  • Appendix 4 Grading Standards for Cell Number and Bleeding Degree in Vitreous
  • Diabetic Retinopathy Early Treatment Research Group (ETDRS) acuity chart measured the best corrected visual acuity (BCVA) of the study eye in the range of 10-73 letters;
  • OCT optical coherence tomography
  • the middle of the eye and the lens are not turbid, and clear three-dimensional fundus photography can be obtained;
  • ADA anti-drug antibody
  • NAb neutralizing antibody
  • PK parameters include but are not limited to: area under the drug-time curve from 0 to time t (AUC0-t), area under the drug-time curve from 0 to infinity (AUC0- ⁇ ), peak concentration (Cmax), time to peak (Tmax) , Clearance (CL), apparent volume of distribution (V), half-life (t1/2), mean residence time (MRT), etc.
  • Continuous variables are described by the number of cases, mean, standard deviation, median, minimum and maximum; categorical variables are described by frequency and percentage.
  • efficacy indicators 95% confidence intervals for the mean of indicators under different dose groups will also be provided.
  • the preliminary effectiveness data of IBI302 in the treatment of nAMD comes from the multi-center, open, dose-escalation study design phase I single-agent dose-escalation study CIBI302A101, with a total of 6 dose groups: 0.05mg/eye, 0.15mg/eye, 0.5mg/eye, 1mg/eye, 2mg/eye, 4mg/eye, eligible subjects received intravitreal injection of study drug on the first day of the study (D1).
  • a 3+3 dose escalation design is adopted, in which the dose group of 0.5 mg/eye to 4 mg/eye must be increased to 6 subjects in the designated dose group before the highest tolerated dose after completing the dose escalation study.
  • a total of 31 nAMD subjects were enrolled and received study drug administration.
  • the average BCVA examination results of the study eyes of subjects in each dose group at each visit were higher than the baseline.
  • the change trend of the inspection result over time and the change of the inspection result from the baseline over time are shown in Figure 19 and Figure 20, respectively.
  • the figure shows the average value of each dose group.
  • OCT Optical Coherence Tomography
  • the average retinal thickness (CST) of 1 mm in the central area measured by OCT was used as an observation index.
  • CST decreased in all dose groups on the 7th day after administration. On the 7th day (i.e.
  • each dose group (0.05mg, 0.15mg, 0.5mg, 1mg, 2mg and 4mg) decreased from baseline by 21.3 ⁇ 29.69 ⁇ m, 8.3 ⁇ 90.47 ⁇ m, 136.8 ⁇ 89.87 ⁇ m, respectively , 142 ⁇ 100.97 ⁇ m, 90.6 ⁇ 41.94 ⁇ m and 117.5 ⁇ 44.70 ⁇ m, the percentages of decrease from baseline are 5%, 2%, 22%, 29%, 17%, 23%, respectively.
  • the 0.5mg and above dose group the degree of CST improvement was significantly higher than the 0.05mg and 0.15mg dose groups.
  • each dose group (0.05mg, 0.15mg, 0.5mg, 1mg, 2mg and 4mg) decreased from baseline by 8.0 ⁇ 67.64 ⁇ m, 23.7 ⁇ 79.53 ⁇ m, 171.8 ⁇ 131.88 ⁇ m, respectively , 203.7 ⁇ 159.66 ⁇ m, 115.3 ⁇ 154.06 ⁇ m and 328.5 ⁇ 191.63 ⁇ m, the percentages of decrease from baseline were 1.7%, 4.4%, 27.4%, 41.1%, 22.0%, 63.3%, respectively.
  • the 0.5mg and above dose group the degree of CST improvement was significantly higher than the 0.05mg and 0.15mg dose groups. The effect of CST improvement continued until the end of the study.
  • CNV choroidal neovascularization
  • the CNV area of the study eye of all subjects decreased by 0.524 ⁇ 3.7806mm 2 on average. Except for the 2 mg dose group, the CNV area of each dose group decreased at 42 days after administration. The CNV area of the contralateral eye decreased by 0.352 ⁇ 1.7891mm 2 on average.
  • the 0.05 mg dose group decreased by 0.980 ⁇ 0.4857 mm 2 from the baseline
  • the 0.15 mg dose group decreased by 4.963 ⁇ 9.9122 mm 2 from the baseline
  • the 0.5 mg dose group decreased by 0.365 ⁇ 1.7140 mm from the baseline. 2.
  • the 1 mg dose group decreased by 0.820 ⁇ 1.7530 mm 2 from the baseline, and the 4 mg dose group decreased by 0.327 ⁇ 0.4206 mm 2 from the baseline.
  • the above data uses the last non-missing value before the first administration of the study drug (including the data during the screening period) as the baseline.
  • Change value measured value-baseline value.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Cell Biology (AREA)
  • Zoology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Ophthalmology & Optometry (AREA)
  • Toxicology (AREA)
  • Epidemiology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

提供了融合蛋白在治疗年龄相关性黄斑变性中的应用。具体而言，提供了抑制补体途径和血管内皮生长因子（VEGF）途径激活的双特异性融合蛋白在治疗年龄相关性黄斑变性中的应用。还提供了包含所述融合蛋白的药物组合物以及使用所述融合蛋白治疗年龄相关性黄斑变性的方法。

Description

融合蛋白在治疗年龄相关性黄斑变性中的应用 技术领域
本发明涉及抑制补体途径和血管内皮生长因子(VEGF)途径激活的双特异性融合蛋白在治疗年龄相关性黄斑变性中的应用。本发明还涉及包含所述融合蛋白的药物组合物以及使用所述融合蛋白治疗年龄相关性黄斑变性的方法。
背景技术
年龄相关性黄斑变性(AMD，又称老年性黄斑变性)是中央区视网膜慢性进展性疾病。老年性黄斑变性大多发生于45岁以上，其患病率随年龄增长而增高，是当前老年人致盲的重要疾病。全球约有3000万老年黄斑变性患者，每年约有50万人因此致盲。随着我国经济发展及人口老龄化的加剧，AMD在我国发病率呈逐年上升的趋势，现已跃居我国第三大致盲原因。
AMD主要表现为视网膜色素上皮细胞(RPE)对视细胞外节盘膜吞噬消化能力下降，结果未能完全消化的盘膜残余小体滞留于基底部细胞原浆中，并向细胞外溢出，沉积于Bruch膜，形成玻璃膜疣。玻璃膜疣处色素上皮、Bruch膜及视细胞发生不同程度的变性、增生或萎缩。美国国家眼科研究所“年龄相关性眼病研究”组将AMD分为早期、中期和晚期AMD，早期AMD以玻璃膜疣为主要特征，发病率高，致盲率低，随着病程进展到晚期，严重影响患者视力，根据其病理特征，可分为萎缩型和渗出型。渗出型AMD也称为湿性AMD，新生血管性AMD，主要病理特征为血管内皮细胞生长因子(VEGF)释放增加，视网膜中心处色素上皮细胞或与光感受器之间未成熟新生血管生成，即脉络膜新生血管生成(CNV)，视网膜下破裂出血，视网膜色素上皮和Bruch膜破坏，发病率低，致盲率高。
AMD的病理机制并没有完全阐明，目前普遍认可VEGF表达增加诱导的血管新生是湿性AMD发病的主要原因。此外，补体基因变异和补体异常活化介导的炎症反应也被认为是AMD发病的重要原因。补体系统的激活对RPE损伤有直接作用，一方面导致RPE萎缩，诱导视网膜中心细胞萎缩，从而形成GA(萎缩型AMD)；另一方面，破坏血-视网膜屏障，导致炎症发生，VEGF高表达，新生血管生成，从而形成新生血管性AMD(渗出型AMD)。
抗补体药物主要用于伴有地图样萎缩的干性AMD，但对于有更高致盲性的湿性AMD疗效并不明确，可能需要与抗VEGF药物联用才能显示优势。但是目前联合治疗采用的是抗补体药物和抗VEGF药物分别玻璃体注射给药治疗，尚无抗补体和抗VEGF的双抗类药物。由于玻璃体注射存在一定的风险，长期重复给药时，患者的顺应性较差，因此这种联合疗法增加了注射操作的风险、给患者心理和经济上都造成了巨大的负担，因此开发能同时抑制VEGF和补体的药物具有潜在的临床价值。此外，补体活化在地图样萎缩的形成中有重要作用，对于合并地图样萎缩的湿性AMD患者，抗补体与抗VEGF二者的融合蛋白可以有重要的临床价值。
WO2013082563A1公开了一种新的抑制VEGF途径和补体途径的融合蛋白。但是，WO2013082563A1中仅验证了该融合蛋白在动物体内的活性，对人类的AMD疾病是否有效 的研究目前还在进行中，而且考虑到人类眼部AMD病症的复杂性、眼部给药在操作和药物容量上的受限性和特殊性等，仍然需要对使用此类融合蛋白的方法和具体方案等进行针对人类AMD疾病的深入研究和改进。
发明内容
一、治疗方法
第一方面，本发明涉及在个体中预防或治疗年龄相关性黄斑变性(AMD)的方法，该方法包括向所述个体施用抑制VEGF途径和补体途径的融合蛋白。
在一个实施方案中，所述融合蛋白包含SEQ ID NO：1的序列：
Figure PCTCN2020138598-appb-000001
在一个优选的实施方案中，所述融合蛋白的序列为SEQ ID NO：1，为方便表述，该融合蛋白在本申请中称为IBI302，其制备方法参见WO2013082563A1。
在一个实施方案中，所述的AMD为湿性AMD。
在一个实施方案中，所述的湿性AMD具有以下的一种或多种症状和体征：视力下降、视物变形、中心暗点、阅读困难、黄斑区网膜肿胀、眼底出血、新生血管形成、瘢痕纤维化形成、地图样萎缩等，或其任意组合。
在一个实施方案中，所述的湿性AMD具有脉络膜新血管形成(choroid neovascularization,CNV)。
在一个实施方案中，所述的湿性AMD为具有以下特征的AMD：
1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
在一个实施方案中，所述个体为人类个体。
在一个优选的实施方案中，所述个体为人类个体，其具有上述实施方案任一项所述的疾病、症状和/或特征。
在一个优选的实施方案中，所述个体为人类个体，其为湿性AMD患者并伴有地图样萎 缩。
在一个优选的实施方案中，所述个体患有具备以下特征的湿性AMD：
1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)中心视网膜厚度较基线降低；
2)脉络膜新生血管生成(CNV)面积降低；
3)最佳矫正视力(BCVA)较基线提高；
4)湿性AMD患者地图样萎缩发生时间滞后；和/或
5)湿性AMD患者地图样萎缩发生时间减慢。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)提高最佳矫正视力(BCVA)；和/或
2)降低中心视网膜厚度；和/或
3)降低脉络膜新生血管生成(CNV)面积。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)最佳矫正视力(BCVA)与基线水平相比提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母；和/或
2)最佳矫正视力(BCVA)与基线水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％，和/或
3)中心视网膜厚度(CST)较基线水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者较基线水平降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％；和/或
4)脉络膜新生血管生成(CNV)面积与基线水平相比降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、 0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与基线水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些实施方案中，本发明的融合蛋白配制为液体药物组合物形式施用。可用的载体和溶剂包括水、林格液、磷酸缓冲盐溶液和等渗氯化钠溶液等。此外，在适宜时也可用无菌、不挥发的油作为溶剂或悬浮介质。为了此目的，可以利用任意混合的非挥发性矿物油或非矿物油，包括合成的甘油单酯或甘油二酯。此外，诸如油酸的脂肪酸也可用于制备注射剂。
在一些实施方案中，包含本发明的融合蛋白的药物组合物为注射用溶液剂或干粉制剂。例如，所述组合物为冻干粉，其可在可药用的液体载体中复配为注射液。所述可药用的液体载体可以是例如无菌的水、林格液、磷酸缓冲盐溶液和等渗氯化钠溶液等。
在一些实施方案中，本发明的融合蛋白通过局部施用。
在一些实施方案中，本发明的融合蛋白通过静脉内注射施用。
在一些实施方案中，本发明的融合蛋白在病灶内施用。
在一些实施方案中，对眼或眼组织直接施用包含本发明的融合蛋白的药物组合物。在一些实施方案中，对眼局部施用药物组合物。在一些实施方案中，通过对眼(眼内注射)或与眼结合的组织注射来施用药物组合物。可以例如通过眼内注射、眼周注射、视网膜下注射、玻璃体内注射、跨隔膜注射、巩膜下注射、脉络膜内注射、前房内注射、结膜下注射、眼球筋膜下注射、眼球后注射、眼球周注射或后巩膜旁递送来施用组合物。还可以例如对个体的玻璃体、视神经、房水、巩膜、结膜、巩膜和结膜之间的区域、视网膜脉络膜组织、黄斑或眼中或靠近眼的其他区域施用组合物。优选地，采用玻璃体内注射给药。
在一些实施方案中，本发明的融合蛋白给药频率为每天三次、每天两次、每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次、每六周一次、每七周一次、每八周一次、或每月一次、每两月一次或更长时间。本发明的融合蛋白的给药周期可以为一周、二周、三周、一月、两月、三月或更长时间，而且每个给药周期之间的间隔可以相同或不同。在一个优选的实施方案中，在每个给药周期给药一次或两次，优选一次，给药方式是每个周期的第一天给药。
在上述的每个实施方案中，本发明的融合蛋白单次给药剂量可选自0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼，例如0.05mg/眼、0.1mg/眼、0.15mg/眼、0.2mg/眼、0.25mg/眼、0.3mg/眼、0.35mg/眼、0.4mg/眼、0.45mg/眼、0.5mg/眼、0.55mg/眼、0.6mg/眼、0.65mg/眼、0.7mg/眼、0.75mg/眼、0.8mg/眼、0.85mg/眼、0.9mg/眼、0.95mg/眼、1mg/眼、1.1mg/眼、1.2mg/眼、1.3mg/眼、1.4mg/眼、1.5mg/眼、1.6mg/眼、1.7mg/眼、1.8mg/眼、1.9mg/眼、2mg/眼、2.1mg/眼、2.2mg/眼、2.3mg/眼、 2.4mg/眼、2.5mg/眼、2.6mg/眼、2.7mg/眼、2.8mg/眼、2.9mg/眼、3mg/眼、3.1mg/眼、3.2mg/眼、3.3mg/眼、3.4mg/眼、3.5mg/眼、3.6mg/眼、3.7mg/眼、3.8mg/眼、3.9mg/眼、4mg/眼、4.1mg/眼、4.2mg/眼、4.3mg/眼、4.4mg/眼、4.5mg/眼、4.6mg/眼、4.7mg/眼、4.8mg/眼、4.9mg/眼、5mg/眼、5.1mg/眼、5.2mg/眼、5.3mg/眼、5.4mg/眼、5.5mg/眼、5.6mg/眼、5.7mg/眼、5.8mg/眼、5.9mg/眼、6mg/眼、6.1mg/眼、6.2mg/眼、6.3mg/眼、6.4mg/眼、6.5mg/眼、6.6mg/眼、6.7mg/眼、6.8mg/眼、6.9mg/眼、7mg/眼、7.1mg/眼、7.2mg/眼、7.3mg/眼、7.4mg/眼、7.5mg/眼、7.6mg/眼、7.7mg/眼、7.8mg/眼、7.9mg/眼、8mg/眼、8.5mg/眼、9mg/眼、9.5mg/眼、10mg/眼，优选2mg/眼或4mg/眼。所述剂量的给药方式优选对眼或眼组织直接施用包含本发明的融合蛋白的药物组合物，例如，对眼局部施用药物组合物，或者，通过对眼(眼内注射)或与眼结合的组织注射来施用药物组合物，例如通过眼内注射、眼周注射、视网膜下注射、玻璃体内注射、跨隔膜注射、巩膜下注射、脉络膜内注射、前房内注射、结膜下注射、眼球筋膜下注射、眼球后注射、眼球周注射或后巩膜旁递送来施用液体组合物。还可以例如对个体的玻璃体、视神经、房水、巩膜、结膜、巩膜和结膜之间的区域、视网膜脉络膜组织、黄斑或眼中或靠近眼的其他区域施用组合物。优选地，采用玻璃体内注射给药。
在一些实施方案中，本发明的融合蛋白或其药物组合物可以单独施用，也可以联合其他药物共同施用。
在上述方法的一些实施方案中，与个体的中心视网膜厚度的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该个体的中心视网膜厚度水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在上述方法的一些实施方案中，与个体的脉络膜新生血管生成(CNV)面积的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该个体的脉络膜新生血管生成(CNV)面积降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在上述方法的一些实施方案中，与个体的最佳矫正视力(BCVA)的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该受试者的最佳矫正视力 (BCVA)提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母，或者与给药前的水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％。
在上述方法的一些实施方案中，在给药后，所述个体不发生严重不良事件。
在上述方法的一些实施方案中，在给药后，所述个体的不良事件发生率与接受安慰剂的个体相当。
在上述方法的一些实施方案中，所述个体是人。
在上述方法的一些具体实施方案中，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼，例如0.05mg/眼、0.1mg/眼、0.15mg/眼、0.2mg/眼、0.25mg/眼、0.3mg/眼、0.35mg/眼、0.4mg/眼、0.45mg/眼、0.5mg/眼、0.55mg/眼、0.6mg/眼、0.65mg/眼、0.7mg/眼、0.75mg/眼、0.8mg/眼、0.85mg/眼、0.9mg/眼、0.95mg/眼、1mg/眼、1.1mg/眼、1.2mg/眼、1.3mg/眼、1.4mg/眼、1.5mg/眼、1.6mg/眼、1.7mg/眼、1.8mg/眼、1.9mg/眼、2mg/眼、2.1mg/眼、2.2mg/眼、2.3mg/眼、2.4mg/眼、2.5mg/眼、2.6mg/眼、2.7mg/眼、2.8mg/眼、2.9mg/眼、3mg/眼、3.1mg/眼、3.2mg/眼、3.3mg/眼、3.4mg/眼、3.5mg/眼、3.6mg/眼、3.7mg/眼、3.8mg/眼、3.9mg/眼、4mg/眼、4.1mg/眼、4.2mg/眼、4.3mg/眼、4.4mg/眼、4.5mg/眼、4.6mg/眼、4.7mg/眼、4.8mg/眼、4.9mg/眼、5mg/眼、5.1mg/眼、5.2mg/眼、5.3mg/眼、5.4mg/眼、5.5mg/眼、5.6mg/眼、5.7mg/眼、5.8mg/眼、5.9mg/眼、6mg/眼、6.1mg/眼、6.2mg/眼、6.3mg/眼、6.4mg/眼、6.5mg/眼、6.6mg/眼、6.7mg/眼、6.8mg/眼、6.9mg/眼、7mg/眼、7.1mg/眼、7.2mg/眼、7.3mg/眼、7.4mg/眼、7.5mg/眼、7.6mg/眼、7.7mg/眼、7.8mg/眼、7.9mg/眼、8mg/眼、8.5mg/眼、9mg/眼、9.5mg/眼、10mg/眼，优选2mg/眼或4mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的中心视网膜厚度水平的给药前水平相比，给药后六周或八周，该个体的中心视网膜厚度水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在上述方法的一些具体实施方案中，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/ 眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼，例如0.05mg/眼、0.1mg/眼、0.15mg/眼、0.2mg/眼、0.25mg/眼、0.3mg/眼、0.35mg/眼、0.4mg/眼、0.45mg/眼、0.5mg/眼、0.55mg/眼、0.6mg/眼、0.65mg/眼、0.7mg/眼、0.75mg/眼、0.8mg/眼、0.85mg/眼、0.9mg/眼、0.95mg/眼、1mg/眼、1.1mg/眼、1.2mg/眼、1.3mg/眼、1.4mg/眼、1.5mg/眼、1.6mg/眼、1.7mg/眼、1.8mg/眼、1.9mg/眼、2mg/眼、2.1mg/眼、2.2mg/眼、2.3mg/眼、2.4mg/眼、2.5mg/眼、2.6mg/眼、2.7mg/眼、2.8mg/眼、2.9mg/眼、3mg/眼、3.1mg/眼、3.2mg/眼、3.3mg/眼、3.4mg/眼、3.5mg/眼、3.6mg/眼、3.7mg/眼、3.8mg/眼、3.9mg/眼、4mg/眼、4.1mg/眼、4.2mg/眼、4.3mg/眼、4.4mg/眼、4.5mg/眼、4.6mg/眼、4.7mg/眼、4.8mg/眼、4.9mg/眼、5mg/眼、5.1mg/眼、5.2mg/眼、5.3mg/眼、5.4mg/眼、5.5mg/眼、5.6mg/眼、5.7mg/眼、5.8mg/眼、5.9mg/眼、6mg/眼、6.1mg/眼、6.2mg/眼、6.3mg/眼、6.4mg/眼、6.5mg/眼、6.6mg/眼、6.7mg/眼、6.8mg/眼、6.9mg/眼、7mg/眼、7.1mg/眼、7.2mg/眼、7.3mg/眼、7.4mg/眼、7.5mg/眼、7.6mg/眼、7.7mg/眼、7.8mg/眼、7.9mg/眼、8mg/眼、8.5mg/眼、9mg/眼、9.5mg/眼、10mg/眼，优选2mg/眼或4mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的脉络膜新生血管生成(CNV)面积的给药前水平相比，给药后六周或八周，该个体的脉络膜新生血管生成(CNV)面积降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在上述方法的一些具体实施方案中，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼，例如0.05mg/眼、0.1mg/眼、0.15mg/眼、0.2mg/眼、0.25mg/眼、0.3mg/眼、0.35mg/眼、0.4mg/眼、0.45mg/眼、0.5mg/眼、0.55mg/眼、0.6mg/眼、0.65mg/眼、0.7mg/眼、0.75mg/眼、0.8mg/眼、0.85mg/眼、0.9mg/眼、0.95mg/眼、1mg/眼、1.1mg/眼、1.2mg/眼、1.3mg/眼、1.4mg/眼、1.5mg/眼、1.6mg/眼、1.7mg/眼、1.8mg/眼、1.9mg/眼、2mg/眼、2.1mg/眼、2.2mg/眼、2.3mg/眼、2.4mg/眼、2.5mg/眼、2.6mg/眼、2.7mg/眼、2.8mg/眼、2.9mg/眼、3mg/眼、3.1mg/眼、3.2mg/眼、3.3mg/眼、3.4mg/眼、3.5mg/眼、3.6mg/眼、3.7mg/眼、3.8mg/眼、3.9mg/眼、4mg/眼、4.1mg/眼、4.2mg/眼、4.3mg/眼、4.4mg/眼、4.5mg/眼、4.6mg/眼、4.7mg/眼、4.8mg/眼、4.9mg/眼、5mg/眼、5.1mg/眼、5.2mg/眼、5.3mg/眼、5.4mg/眼、5.5mg/眼、5.6mg/眼、5.7mg/眼、5.8mg/眼、5.9mg/眼、6mg/眼、6.1mg/眼、6.2mg/眼、6.3mg/眼、6.4mg/眼、6.5mg/眼、6.6mg/眼、6.7mg/眼、6.8mg/眼、6.9mg/眼、7mg/眼、7.1mg/眼、7.2mg/眼、7.3mg/眼、7.4mg/眼、7.5mg/眼、7.6mg/眼、7.7mg/眼、7.8mg/眼、7.9mg/眼、8mg/眼、8.5mg/眼、9mg/眼、9.5mg/眼、10mg/眼，优选2mg/眼或4mg/眼的单次剂量，每 天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的最佳矫正视力(BCVA)的给药前水平相比，给药后六周或八周，该个体的最佳矫正视力(BCVA)提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母，或者与给药前的水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％。
二、用途
第二方面，本发明涉及抑制VEGF途径和补体途径的融合蛋白在制备用于在个体中预防或治疗年龄相关性黄斑变性(AMD)的药物中的用途。
第三方面，本发明涉及抑制VEGF途径和补体途径的融合蛋白，其用于在个体中预防或治疗年龄相关性黄斑变性(AMD)。
对于第二和第三方面，对它们的进一步限定如下：
在一个实施方案中，所述融合蛋白包含SEQ ID NO：1的序列：
Figure PCTCN2020138598-appb-000002
在一个优选的实施方案中，所述融合蛋白的序列为SEQ ID NO：1，为方便表述，该融合蛋白在本申请中称为IBI302，其制备方法参见WO2013082563A1。
在一个实施方案中，所述的AMD为湿性AMD。
在一个实施方案中，所述的湿性AMD具有以下的一种或多种症状和体征：视力下降、视物变形、中心暗点、阅读困难、黄斑区网膜肿胀、眼底出血、新生血管形成、瘢痕纤维化形成、地图样萎缩，或其任意组合。
在一个实施方案中，所述的湿性AMD具有脉络膜新血管形成(CNV)。
在一个实施方案中，所述的湿性AMD为具有以下特征的AMD：
1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
在一个实施方案中，所述个体为人类个体。
在一个优选的实施方案中，所述个体为人类个体，其具有上述实施方案任一项所述的疾病、症状和/或特征。
在一个优选的实施方案中，所述个体为人类个体，其为湿性AMD患者并伴有地图样萎缩。
在一个优选的实施方案中，所述个体患有具备以下特征的湿性AMD：
1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)中心视网膜厚度较基线降低；
2)脉络膜新生血管生成(CNV)面积降低；
3)最佳矫正视力(BCVA)较基线提高；
4)湿性AMD患者地图样萎缩发生时间滞后；和/或
5)湿性AMD患者地图样萎缩发生时间减慢。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)提高最佳矫正视力(BCVA)；
2)降低中心视网膜厚度；和/或
3)降低脉络膜新生血管生成(CNV)面积。
在一个优选的实施方案中，所述治疗可在个体中实现以下一种或多种效果：
1)最佳矫正视力(BCVA)与基线水平相比提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母；和/或
2)最佳矫正视力(BCVA)与基线水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、 70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％，和/或
3)中心视网膜厚度(CST)较基线水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者较基线水平降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％；和/或
4)脉络膜新生血管生成(CNV)面积与基线水平相比降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与基线水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些实施方案中，本发明的融合蛋白给药频率为每天三次、每天两次、每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次、每六周一次、每七周一次、每八周一次、或每月一次、每两月一次或更长时间。本发明的融合蛋白的给药周期可以为一周、二周、三周、一月、两月、三月或更长时间，而且每个给药周期的间隔可以相同或不同。在一个优选的实施方案中，在每个给药周期给药一次或两次，优选一次，给药方式是每个周期的第一天给药。
在上述的每个实施方案中，本发明的融合蛋白单次给药剂量选自0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼，例如0.05mg/眼、0.1mg/眼、0.15mg/眼、0.2mg/眼、0.25mg/眼、0.3mg/眼、0.35mg/眼、0.4mg/眼、0.45mg/眼、0.5mg/眼、0.55mg/眼、0.6mg/眼、0.65mg/眼、0.7mg/眼、0.75mg/眼、0.8mg/眼、0.85mg/眼、0.9mg/眼、0.95mg/眼、1mg/眼、1.1mg/眼、1.2mg/眼、1.3mg/眼、1.4mg/眼、1.5mg/眼、1.6mg/眼、1.7mg/眼、1.8mg/眼、1.9mg/眼、2mg/眼、2.1mg/眼、2.2mg/眼、2.3mg/眼、2.4mg/眼、2.5mg/眼、2.6mg/眼、2.7mg/眼、2.8mg/眼、2.9mg/眼、3mg/眼、3.1mg/眼、3.2mg/眼、3.3mg/眼、3.4mg/眼、3.5mg/眼、3.6mg/眼、3.7mg/眼、3.8mg/眼、3.9mg/眼、4mg/眼、4.1mg/眼、4.2mg/眼、4.3mg/眼、4.4mg/眼、4.5mg/眼、4.6mg/眼、4.7mg/眼、4.8mg/眼、4.9mg/眼、5mg/眼、5.1mg/眼、5.2mg/眼、5.3mg/眼、5.4mg/眼、5.5mg/眼、5.6mg/眼、5.7mg/眼、5.8mg/眼、5.9mg/眼、6mg/眼、6.1mg/眼、6.2mg/眼、6.3mg/眼、6.4mg/眼、6.5mg/眼、6.6mg/眼、6.7mg/眼、6.8mg/眼、6.9mg/眼、7mg/眼、7.1mg/眼、7.2mg/眼、7.3mg/眼、7.4mg/眼、7.5mg/眼、7.6mg/眼、7.7mg/眼、7.8mg/眼、7.9mg/眼、8mg/眼、8.5mg/眼、9mg/眼、9.5mg/眼、10mg/眼，优选2mg/眼或4mg/眼。
在一些实施方案中，本发明的融合蛋白或其药物组合物可以单独施用，也可以联合其他 药物共同施用。
在一些实施方案中，与个体的中心视网膜厚度的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该个体的中心视网膜厚度水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些实施方案中，与个体的脉络膜新生血管生成(CNV)面积的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该个体的脉络膜新生血管生成(CNV)面积降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些实施方案中，与个体的最佳矫正视力(BCVA)的给药前水平相比，在给药后2周、3周或4周、给药后5周、给药后6周、给药后7周、给药后8周、给药后9周、给药后10周、给药后3个月、给药后4个月或给药后5个月，该受试者的最佳矫正视力(BCVA)提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母，或者与给药前的水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％。
在一些实施方案中，在给药后，所述个体不发生严重不良事件。
在一些实施方案中，在给药后，所述个体的不良事件发生率与接受安慰剂的个体相当。
在一些实施方案中，所述个体是人。
在一些具体实施方案中，所述融合蛋白以下述方案向所述个体施用：以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼的单次剂量，每天一 次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的中心视网膜厚度水平的给药前水平相比，给药后六周或八周，该个体的中心视网膜厚度水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些具体实施方案中，所述融合蛋白以下述方案向所述个体施用：以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的脉络膜新生血管生成(CNV)面积的给药前水平相比，给药后六周或八周，该个体的脉络膜新生血管生成(CNV)面积降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
在一些具体实施方案中，所述融合蛋白以下述方案向所述个体施用：以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼，例如0.5-2mg/眼、1-3mg/眼、2-4mg/眼、2-3mg/眼或3-5mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的最佳矫正视力(BCVA)的给药前水平相比，给药后六周或八周，该个体的最佳矫正视力(BCVA)提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母，或者与给药前的水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％。
应当理解，上文第二至三方面记载的任何技术特征与第一方面中记载的技术方案的任何技术特征的任意组合得到的技术方案也包括在本申请中。
三、单次药物剂量单元或成套药物包装盒
在第四方面，本发明涉及一种单次药物剂量单元，其特征在于包含抑制VEGF途径和补体途径的融合蛋白。
在一个实施方案中，所述融合蛋白包含SEQ ID NO：1的序列：
Figure PCTCN2020138598-appb-000003
在一个优选的实施方案中，所述融合蛋白的序列为SEQ ID NO：1，为方便表述，该融合蛋白在本申请中称为IBI302，其制备方法参见WO2013082563A1。
在一些实施方案中，所述单次药物剂量单元包含以下剂量的所述融合蛋白：0.01-10mg，优选0.05-8mg，更优选0.05-6mg，更优选0.5-5mg，更优选1-5mg，更优选2-5mg，例如0.5-2mg、1-3mg、2-4mg、2-3mg或3-5mg，例如0.05mg、0.1mg、0.15mg、0.2mg、0.25mg、0.3mg、0.35mg、0.4mg、0.45mg、0.5mg、0.55mg、0.6mg、0.65mg、0.7mg、0.75mg、0.8mg、0.85mg、0.9mg、0.95mg、1mg、1.1mg、1.2mg、1.3mg、1.4mg、1.5mg、1.6mg、1.7mg、1.8mg、1.9mg、2mg、2.1mg、2.2mg、2.3mg、2.4mg、2.5mg、2.6mg、2.7mg、2.8mg、2.9mg、3mg、3.1mg、3.2mg、3.3mg、3.4mg、3.5mg、3.6mg、3.7mg、3.8mg、3.9mg、4mg、4.1mg、4.2mg、4.3mg、4.4mg、4.5mg、4.6mg、4.7mg、4.8mg、4.9mg、5mg、5.1mg、5.2mg、5.3mg、5.4mg、5.5mg、5.6mg、5.7mg、5.8mg、5.9mg、6mg、6.1mg、6.2mg、6.3mg、6.4mg、6.5mg、6.6mg、6.7mg、6.8mg、6.9mg、7mg、7.1mg、7.2mg、7.3mg、7.4mg、7.5mg、7.6mg、7.7mg、7.8mg、7.9mg、8mg、8.5mg、9mg、9.5mg、10mg，优选2mg或4mg。
在第五方面，本发明涉及一种成套药物包装盒，其特征在于，包含上述单次药物剂量单元实施方案中任意一项所述的融合蛋白。
在第六方面，本发明涉及如上所述的单次药物剂量单元或如上所述的成套药盒在制备预防或治疗年龄相关性黄斑变性(AMD)的药物中的用途。
在第七方面，本发明涉及如上所述的单次药物剂量单元或如上所述的成套药盒，其用于预防或治疗年龄相关性黄斑变性(AMD)。
应当理解，上文第四至七方面记载的任何技术特征与第一至三方面中记载的技术方案的任何技术特征的任意组合得到的技术方案也包括在本申请中。
定义
本文所使用的术语具有如下的定义。若本文没有定义，可适用专利申请WO2013082563A1中的定义。如果在两者中都没有给出定义，则本发明中所使用的术语具有本领域所通常理解的含义。
为了解释本说明书，将使用以下定义，并且只要适当，以单数形式使用的术语也可以包括复数，并且反之亦然。要理解，本文所用的术语仅是为了描述具体的实施方案，并且不意欲是限制性的。
如本文所用，术语“和/或”是指可选项中的任一项或可选项的两项或更多项。
如本文所用，术语“包含”或“包括”是指包括所述的要素、整数或步骤，但是不排除任意其他要素、整数或步骤。在本文中，当使用术语“包含”或“包括”时，除非另有指明，否则也涵盖由所述及的要素、整数或步骤组合的情形。例如，当提及“包含”某个具体序列的抗体可变区时，也旨在涵盖由该具体序列组成的抗体可变区。
“个体”包括哺乳动物。哺乳动物包括但不限于，家养动物(例如，牛，羊，猫，狗和马)，灵长类动物(例如，人和非人灵长类动物如猴)，兔，以及啮齿类动物(例如，小鼠和大鼠)。在一些实施方案中，个体是人，包括儿童、青少年或成人。
术语“组合施用”是指施用两种或更多种治疗剂以治疗如本公开所述的疾病。这种施用包括以基本上同时的方式共同施用这些治疗剂，例如以具有固定比例的活性成分的单一组合物。或者，这种施用包括对于各个活性成分在多种或在分开的容器(例如片剂、胶囊、粉末和液体)中的共同施用。粉末和/或液体可以在施用前重构或稀释至所需剂量。此外，这种施用还包括以大致相同的时间或在不同的时间以顺序的方式使用每种类型的治疗剂。在任一情况下，治疗方案将提供药物组合在治疗本文所述的病症或病状中的有益作用。
用于本文时，“治疗”指减缓、中断、阻滞、缓解、停止、降低、或逆转已存在的症状、病症、病况或疾病的进展或严重性。
用于本文时，“预防”包括对疾病或病症或特定疾病或病症的症状的发生或发展的抑制。在一些实施方式中，具有家族病史的个体是预防性方案的候选。通常，术语“预防”是指在病征或症状发生前，特别是在具有风险的个体中发生前的药物施用。
术语“有效量”指本发明的制剂或融合蛋白的量或剂量，其以单一或多次剂量施用患者后，在治疗的患者中产生预期效果。有效量可以由作为本领域技术人员的主治医师通过考虑以下多种因素来容易地确定：诸如哺乳动物的物种；它的大小、年龄和一般健康；涉及的具体疾病；疾病的程度或严重性；个体患者的应答；施用的具体抗体；施用模式；施用制剂的生物利用率特征；选择的给药方案；和任何伴随疗法的使用。
“治疗有效量”指以需要的剂量并持续需要的时间段，有效实现所需治疗结果的量。本发明的制剂、抗体或抗体片段或其缀合物或组合物的治疗有效量可以根据多种因素如疾病状态、个体的年龄、性别和重量和抗体或抗体部分在个体中激发所需反应的能力而变动。治疗有效量也是这样的一个量，其中制剂、抗体或抗体片段或其缀合物或组合物的任何有毒或有害作用不及治疗有益作用。
“预防有效量”指以需要的剂量并持续需要的时间段，有效实现所需预防结果的量。通常，由于预防性剂量在对象中在疾病较早阶段之前或在疾病较早阶段使用，故预防有效量将小于 治疗有效量。
术语“融合蛋白”指具有共价连接在一起的两个或多个部分的多肽，其中所述部分中的每一个源自不同的蛋白质。该两个或多个部分可以直接通过单个肽键或通过含有一个或多个氨基酸残基的肽接头连接。通常，该两个部分和该接头将彼此处于可读框中，并用重组技术产生。
术语“抑制VEGF途径和补体途径的融合蛋白”包含补体抑制结构域(CID)、VEGF抑制结构域(VID)和半寿期延长结构域，在本发明一个优选地实施方案中，所述的VEGF融合蛋白选自WO2013082563A1中的SEQ ID NO:40，所述“CID”、“VID”及“半衰期延长结构域参见WO2013082563A1。
术语“中心视网膜”，位于视网膜中央，是视力最敏感区，负责明视觉和色觉的视锥细胞就分布于该区域，因此任何累及该区域的病变都会引起中心视力的明显下降，亦可合并色觉异常、视物变形、中心暗点等，该部病变可表现为出血、水肿、厚度增加以及萎缩等。另外，黄斑位于中心视网膜区域的最中心。
术语“脉络膜新生血管”，是指来自脉络膜毛细血管的增殖血管，通过Bruch膜的裂口而扩展，在Bruch膜与视网膜色素上皮之间、或神经视网膜与视网膜色素上皮之间、或位于视网膜色素上皮与脉络膜之间增殖形成，许多累及RPE-Bruch膜-脉络膜毛细血管复合体的疾病均可导致脉络膜新生血管的形成。
术语“最佳矫正视力”，即完全矫正眼球的屈光不正后测得的视力。代表了眼球实际的视物能力。
如文中所用，术语“制剂”或“药物组合物”指适合于向动物优选哺乳动物(包括人)施用的包含至少一种活性成分和至少一种非活性成分的组合物。“液体制剂”或“液体组合物”是指液体形式的制剂。本发明的液体组合物包含(i)本发明所述的融合蛋白；(ii)缓冲剂；和(iii)溶媒。本发明的制剂的组成可如上面涉及液体药物组合物的实施方案中所示。本发明的液体制剂优选为注射剂。
“可药用载体”指药物制剂中除活性成分以外的成分，其对对象无毒性。可药用载体包括但不限于缓冲剂、赋形剂、稳定剂或防腐剂。
如文中所用，“缓冲剂”是指pH缓冲剂。例如，缓冲剂选自组氨酸、谷氨酸盐、磷酸盐、乙酸盐、柠檬酸盐和三羟甲基氨基甲烷。
如文中所用，术语“溶媒”是指用于溶解或悬浮活性成分和非活性成分以形成液体制剂的液体。可用于本发明的溶媒包括但不限于注射用水、注射用有机溶剂包括但不限于注射用油、乙醇、丙二醇等，或其组合。
如文中所用，术语“单次药物剂量单元”是指在给药时待施用于患者的包含本发明的融合蛋白的单次药物剂型，例如注射用的小瓶、安瓿、预充针或预充式注射器，其中含有药物的溶液或冻干粉。
术语“约”在与数字数值联合使用时意为涵盖具有比指定数字数值小10％的下限和比指定数字数值大10％的上限的范围内的数字数值。
附图说明
图1描述了IBI302对VEGF-A诱导HUVEC增殖的抑制作用。
图2描述了IBI302对补体经典途径介导溶血的抑制作用。
图3描述了IBI302对补体旁路途径介导溶血的抑制作用。
图4描述了各剂量IBI302对CNV的抑制作用。*P＜0.05vs PBS组；#P＜0.05vs IBI302 1、3或5μg组。
图5描述了IBI302抑制CNV的形成和渗漏。*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
图6描述了IBI302抑制炎性细胞聚集的作用。*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
图7描述了IBI302对恒河猴CNV模型眼底荧光渗漏的影响(荧光造影)。
图8描述了脉络膜CD31免疫组化染色代表图。
图9描述了脉络膜C5b-9免疫组化染色代表图。
图10描述了恒河猴静脉注射及玻璃体内注射IBI302后血清中浓度药-时曲线。
图11描述了恒河猴玻璃体内注射IBI302后房水和玻璃体液中浓度药-时曲线。
图12描述了恒河猴玻璃体内注射IBI302后晶状体、角膜、虹膜、视网膜、脉络膜、视神经、巩膜中浓度药-时曲线。
图13描述了恒河猴首次静脉注射及玻璃体内注射不同剂量的IBI302后平均血药浓度-时间变化曲线。
图14描述了恒河猴末次静脉注射及玻璃体内注射不同剂量的IBI302后平均血药浓度-时间变化曲线。
图15描述了实施例5所述的I期临床研究的剂量递增示意图。
图16描述了实施例5所述的I期临床研究的研究流程示意图。
图17描述了实施例5所述的I期临床研究的初期试验结果显示的正确识别字母数变化趋势及与基线比较。
图18描述了实施例5所述的I期临床研究的初期试验结果显示的中心视网膜厚度(CST)变化趋势以及与基线比较(△CST)。
图19描述了实施例5所述的I期临床研究的总体研究结果中的各剂量组BCVA(EDTRS字母数)检查结果随时间的变化，其中采用首次给予研究药物前(包括筛选期的数据)的最后一次非缺失值作为基线。
图20描述了实施例5所述的I期临床研究的总体研究结果中的各剂量组BCVA(EDTRS字母数)与基线相比的变化值随时间的变化，其中采用首次给予研究药物前(包括筛选期的数据)的最后一次非缺失值作为基线。变化值定义为各访视测量值与基线差值的平均值。
图21描述了实施例5所述的I期临床研究的总体研究结果中的各剂量组眼科OCT检查黄斑中心区域厚度随时间的变化，其中采用首次给予研究药物前(包括筛选期的数据)的最后一次非缺失值作为基线。
图22描述了实施例5所述的I期临床研究的总体研究结果中的各剂量组眼科OCT检查黄斑中心区域厚度与基线相比的变化值随时间的变化，其中采用首次给予研究药物前(包括筛选期的数据)的最后一次非缺失值作为基线。变化值定义为各访视测量值与基线差值的平均值。
具体实施方式
以下结合实施例进一步说明本发明，下列实施例不应被理解为对本发明的限制。
缩写
AMD   年龄相关性黄斑变性
BCVA  最佳矫正视力
OCT   光学相干断层成像
CNV   脉络膜新生血管生成
AE    不良反应
DLT   剂量限制性毒性
EC50  半数有效浓度
SAE   严重不良反应
TEAE  治疗期间不良反应
CR1   人1型补体受体
实施例1：体外药效学研究
IBI302与VEGF家族和补体家族细胞因子的亲和力
采用Biacore T200考察IBI302与相关配体的亲和力，IBI302与VEGF家族和C3b、C4b的亲和力，见表1。
IBI302与VEGF家族细胞因子的亲和力与阿柏西普注射液相当，显著强于贝伐珠单抗注射液。IBI302与C4b的亲和力和CR1相当；IBI302与C3b的亲和力略弱于CR1，可能是因为IBI302的CID端只是CR1的部分结构域所致，但是仍然达到nM级。
表1.IBI302和三种对照品与VEGF家族或者补体家族细胞因子的亲和力
Figure PCTCN2020138598-appb-000004
NB：未检测到。
IBI302对VEGF-A诱导HUVEC增殖的抑制作用
以一系列浓度梯度的IBI302与固定浓度的VEGF-A165作用后，HUVEC细胞随着游离的VEGF-A165的量不同而表现出不同的增殖能力，利用CCK-8显色试剂盒测定HUVEC活细胞数量，从而测定IBI302对HUVEC细胞增殖抑制效率。同时以阿柏西普和贝伐珠单抗注射液为对照品。
结果显示：IBI302的半数有效浓度(EC50)为54.36ng/ml和57.53ng/ml，相当于347pmol/L和368pmol/L；阿柏西普注射液的EC50为41.02ng/ml，相当于357pmol/L；贝伐珠单抗注射液的EC50为87.07ng/ml，相当于584pmol/L。IBI302能显著抑制VEGF刺激的HUVEC增殖，与阿柏西普的活性相似，比贝伐珠单抗注射液稍强。见表2和图1。
表2.IBI302对VEGF-A诱导HUVEC增殖的抑制作用
Figure PCTCN2020138598-appb-000005
*由于采用两块板进行试验，因此IBI302做了2次。
IBI302对补体经典途径的抑制作用
采用人血清补体刺激致敏绵羊红细胞模型，IBI302在0.02～1080.26nM浓度范围内能够浓度依赖性的抑制人血清补体诱导的致敏绵羊红细胞的溶血反应，半数有效浓度(EC50)值为2.514nM。CR1的EC50值为2.254nM。提示IBI302能显著抑制补体经典途径，与CR1相似。具体结果见图2。
IBI302对补体旁路途径的抑制作用
采用人血清补体刺激兔红细胞模型，IBI302在1.50～383.39nM浓度范围内能够浓度依赖性的抑制人血清补体诱导的兔红细胞的溶血反应，EC50值为14.59nM。CR1的EC50值为15.30nM。提示IBI302能显著抑制补体旁路途径，与CR1相似。具体结果见图3。
IBI302对补体激活的氧化损伤的hRPE细胞屏障功能的作用及其机制
实验采用hRPE细胞，分为正常组(nor)、模型组(ctr)、阿柏西普组(VEGF-Trap)、CR1组、IBI302组。除正常组外各组均加入t-BHP和10％正常人血清，正常组加入等体积的PBS。药物治疗组再分别加入相应药物，并使各药物终浓度为1μg/ml，正常组和模型组加入等体积的PBS。培养4h后检测hRPE单层细胞的屏障功能。培养4h后ELISA方法检测hRPE细胞分泌的VEGF、趋化因子配体2(Chemokine(C-C motif)ligand 2，CCL2)、C3a、C5a和MAC。
模型组(ctr)、阿柏西普组，CR1组及IBI302组补体激活的氧化损伤hRPE的跨上皮电阻值(TER)值分别为正常组(nor)hRPE的54.01％，64.64％，63.48％及75.90％。阿柏西普组、CR1组和IBI302组与模型组比较TER均显著提高，IBI302组与阿柏西普组，CR1组比较亦有显著差异，见表3。
表3.IBI302对跨上皮电阻值的影响
Figure PCTCN2020138598-appb-000006
Figure PCTCN2020138598-appb-000007
*P＜0.05vs模型组；#P＜0.05vs IBI302组。
与模型组相比，阿柏西普组、CR1组和IBI302组hRPE分泌VEGF的浓度明显降低。IBI302组与阿柏西普组和CR1组相比，VEGF浓度明显降低。CR1组和IBI302组hRPE分泌CCL2较模型组明显降低；而阿柏西普组hRPE分泌的CLL2与模型组相比统计学差异不显著。IBI302组与阿柏西普组和CR1组相比，CCL2浓度明显降低，见表4和表5。
表4.IBI302对VEGF表达的影响
Figure PCTCN2020138598-appb-000008
*P＜0.05vs模型组；#P＜0.05vs IBI302组。
表5.IBI302对CCL2表达的影响
Figure PCTCN2020138598-appb-000009
*P＜0.05vs模型组；#P＜0.05vs IBI302组。
对hRPE补体激活影响的实验结果表明：与模型组(ctr)相比，CR1组和IBI302组均明显降低补体活性成分C3a、C5a、MAC的表达。与CR1组相比，IBI302组抑制C3a、C5a和MAC的表达无统计学差异。而阿柏西普组与模型组比较对补体活性成分C3a、C5a、MAC生成的影响无统计学差异，见表6、7、8。
表6.IBI302对C3a表达的影响
Figure PCTCN2020138598-appb-000010
*P＜0.05vs模型组。
表7.IBI302对C5a表达的影响
Figure PCTCN2020138598-appb-000011
Figure PCTCN2020138598-appb-000012
*P＜0.05vs模型组。
表8.IBI302对MAC形成的影响
Figure PCTCN2020138598-appb-000013
*P＜0.05vs模型组。
这些实验结果表明IBI302对补体激活的氧化损伤的hRPE细胞屏障功能具有保护作用，其机制可能是通过同时抑制VEGF和补体激活后的补体活性成分C3a、C5a以及MAC的表达，进一步减少hRPE分泌的CCL2和VEGF。
实施例2：动物体内药效学研究
IBI302对激光诱导的小鼠脉络膜新生血管的作用及机制研究
实验采用C57BL/6J小鼠，激光光凝4个点，随机将纳入研究对象的小鼠随机分为6组，每组2只(4眼)小鼠：激光造模组，磷酸盐缓冲液(PBS)治疗组，IBI302 1μg/μl、3μg/μl、5μg/μl、10μg/μl剂量组。激光光凝后立即向各组脉络膜新生血管生成(CNV)小鼠的双眼玻璃体腔中各注射1μl各剂量的IBI302或者PBS。经过激光光凝后第7天异硫氰酸荧光素-右旋糖酐(FITC-Dextran)眶静脉窦灌注和脉络膜铺片，观察各剂量IBI302对激光诱导的小鼠CNV面积的影响。
进一步实验分组为：IBI302组，阿柏西普(VEGF-Trap)组，CR1组，PBS组。IBI302和对照药物均为10μg/μl，激光光凝后立即向各组CNV小鼠的双眼玻璃体腔中各注射1μl各组样品。激光光凝后第7天小鼠进行眼底造影检查，观察CNV的情况。小鼠采用水合氯醛麻醉后用复方托品卡胺散瞳。首先拍摄红外眼底照片，然后分别进行FITC-Dextran、吲哚青绿血管造影(ICGA)和眼底荧光血管造影(FFA)眼底造影检查。最后用ELISA法检测RPE-脉络膜中VEGF、CCL2、TNF-α以及C3a、C5a蛋白浓度。
实验结果：
不同剂量IBI302对CNV的抑制作用：激光光凝后第7天行小鼠眶静脉窦FITC-Dextran灌注后脉络膜铺片，见新生血管呈花瓣状结构。激光造模组(laser组)与PBS组的CNV面积无统计学差异。与PBS组相比，IBI302 1μg组、3μg组、5μg组、10μg组CNV面积分别比PBS组减少14.27％，48.79％，62.46％，68.83％。IBI302 10μg组与1μg组、3μg组、5μg组之间两两比较的CNV面积差别均有统计学意义，见表9和图4。
表9.各剂量IBI302组眶静脉窦灌注FITC-Dextran后CNV面积
Figure PCTCN2020138598-appb-000014
Figure PCTCN2020138598-appb-000015
*P＜0.05vs PBS组；#P＜0.05vs IBI302 1、3或者5μg组。
与PBS对照组相比，10μg/眼IBI302、阿柏西普与CR1均抑制CNV的面积和渗漏。与阿柏西普和CR1组相比，10μg/眼IBI302能明显抑制CNV面积和渗漏，见表10、11和图5。
表10.FFA中各组渗漏面积百分比
Figure PCTCN2020138598-appb-000016
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
表11.ICGA中各组CNV面积百分比
Figure PCTCN2020138598-appb-000017
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组
IBI302对激光诱导的CNV小鼠VEGF生成的抑制作用：激光光凝后第3天，与PBS组相比，10μg/眼IBI302、阿柏西普和CR1均可抑制激光诱导的CNV小鼠VEGF的生成。与阿柏西普组和CR1组相比，IBI302抑制VEGF生成的作用的差异有统计学意义，见表12。
表12.IBI302对VEGF表达的影响
Figure PCTCN2020138598-appb-000018
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
IBI302抑制激光诱导的CNV小鼠补体激活：激光光凝后第3天，脉络膜免疫荧光铺片发现与PBS组、阿柏西普组、CR1组相比，10μg/眼IBI302可明显抑制MAC在脉络膜的沉积。激光光凝后12h，ELISA法检测RPE-脉络膜复合物中C3a和C5a的表达水平发现，与模型组相比，CR1组和IBI302组中C3a和C5a的表达明显降低。而IBI302组与CR1组间C3a与C5a的浓度无统计学差异；IBI302组与阿柏西普组相比，C3a与C5a的浓度明显降低，见表13和表14。
表13.IBI302对C3a表达的影响
Figure PCTCN2020138598-appb-000019
Figure PCTCN2020138598-appb-000020
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
表14.IBI302对C5a表达的影响
Figure PCTCN2020138598-appb-000021
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
IBI302对激光诱导的CNV小鼠炎性因子CCL2和TNF-α的抑制作用：分别在激光光凝后第12h和第3天用ELISA检测RPE-脉络膜复合物中炎性因子CCL2和TNF-α的表达水平。与PBS组相比，10μg/眼CR1组和IBI302组中炎性因子CCL2和TNF-α的表达明显降低。CCL2和TNF-α浓度在IBI302组显著低于阿柏西普组，见表15和表16。
表15.IBI302对CCL2表达的影响
Figure PCTCN2020138598-appb-000022
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
表16.IBI302对TNF-α表达的影响
Figure PCTCN2020138598-appb-000023
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
IBI302对激光诱导的CNV小鼠炎性细胞聚集的抑制作用：激光光凝后第1天和第3天分别脉络膜铺片免疫荧光检测中性粒细胞和巨噬细胞在脉络膜的聚集。IBI302可明显抑制中性粒细胞和巨噬细胞在脉络膜的沉积，与PBS组和阿柏西普组相比均有统计学差异，见表17、表18和图6。
表17.IBI302对中性粒细胞聚集的影响
Figure PCTCN2020138598-appb-000024
Figure PCTCN2020138598-appb-000025
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组
表18.IBI302对巨噬细胞聚集的影响
Figure PCTCN2020138598-appb-000026
*P＜0.05vs PBS组；#P＜0.05vs IBI302 10μg组。
结果表明IBI302可抑制激光诱导的小鼠CNV的形成和渗漏，其可能机制是通过同时抑制VEGF和补体激活后活性成分C3a、C5a、MAC和CCL2、TNF-α的表达，抑制巨噬细胞和中性粒细胞的浸润，从而抑制VEGF分泌和新生血管生成。
IBI302对激光致恒河猴脉络膜新生血管抑制试验
实验采用激光围绕恒河猴眼底黄斑中心凹光凝，诱导眼底脉络膜血管新生，建立与人类脉络膜新生血管类似的动物模型。选择双眼造模成功的25只猴(雌雄兼有)分5组，分别为模型对照组、贝伐珠单抗注射液1.25mg/眼组及IBI302 0.25mg/眼组、IBI302 0.5mg/眼组、IBI302 1.25mg/眼组，每组5只猴，雌雄兼有。具体见表19。光凝后21天，各组猴按50μl/眼经玻璃体单次注射相应药物。
给药后14、28天进行眼底彩色照相、荧光素眼底血管造影，11、25天进行光学相干断层扫描(OCT)检查观察供试品对脉络膜新生血管的抑制情况。29天对猴双眼抽取房水进行VEGF检测，取左眼进行组织病理学检查，右眼进行CD31、C5b-9免疫组织化学染色检查。
表19.分组和给药设计表
Figure PCTCN2020138598-appb-000027
结果显示：
荧光造影和OCT检查：模型对照组猴给药后14、28天荧光素渗漏程度均未见明显变化，荧光斑面积亦未见明显缩小，部分猴眼荧光素渗漏甚至可见随时间进程加重的趋势。4级荧光斑数与给药前相比未见明显变化。给药后11、25天OCT检查视网膜厚度未见明显降低，局部Bruch’s膜结构未见修复，脉络膜新生血管仍明显存在，可见高反射光团。
贝伐珠单抗注射液组给药后14、28天渗漏面积较给药前明显缩小，与模型对照组相比有显著差异(P＜0.05)，4级荧光斑数与给药前相比明显减少。给药后11、25天OCT检查所有眼球病损最高处视网膜厚度均明显降低，甚至接近或达到造模前水平，与对照组相比差异均有统计学意义(P＜0.05)。
IBI302 0.25mg/眼组、0.5mg/眼组和1.25mg/眼组猴给药后14、28天渗漏面积较给药前明显缩小，与模型对照组相比有显著差异(P＜0.05)，且给药后28天的渗漏面积改善也显著高于阳性对照组(P＜0.05)。4级荧光斑数与给药前相比明显减少。给药后11、25天OCT检查视网膜厚度均明显降低，甚至接近或达到造模前水平，与模型对照组相比差异均有统计学意义(P＜0.05)，与阳性对照组相比，各指标差异无统计学意义(P＞0.05)。
给药前后各组猴眼底激光斑荧光素渗漏、荧光光斑数和视网膜厚度分别见表20、表21、表22和图7。
表20.IBI302对恒河猴荧光素渗漏面积减少量(mm 2)及改善率的影响
Figure PCTCN2020138598-appb-000028
*与模型对照组比较，P≤0.05；#与阳性对照组比较，P≤0.05。
表21.IBI302对恒河猴荧光斑等级的影响
Figure PCTCN2020138598-appb-000029
*与模型对照组比较，P≤0.05。
表22.IBI302对恒河猴视网膜厚度减少量及改善率的影响
Figure PCTCN2020138598-appb-000030
*与模型对照组比较，P≤0.05。
VEGF检测：给药后14和28天，贝伐珠单抗注射液1.25mg/眼组和模型对照组分别有1 只猴血清VEGF浓度测得在定量下限附近的数值外，其余时间点均低于定量下限。给药后29天，贝伐珠单抗注射液1.25mg/眼组猴房水VEGF浓度与对照组相比降低，差异有统计学意义(P＜0.05)，IBI302各组猴房水VEGF浓度均低于定量下限。具体结果见表23。
表23.IBI302对恒河猴血清及房水VEGF指标的影响(pg/ml)
Figure PCTCN2020138598-appb-000031
-BLOQ(低于检测下限)无法进行统计；*与模型对照组比较，P≤0.05。
组织病理学与免疫组化：HE染色结果显示：模型对照组出现轻微～中度的灶性视网膜/脉络膜纤维组织增生和轻微～轻度的灶性神经上皮脱离。IBI302各组及贝伐珠单抗注射液1.25mg/眼组上述病变严重程度均较模型对照组轻。
Masson三色染色结果显示：模型对照组出现轻微视网膜内灶性胶原纤维增生阳性。与模型对照组相比，贝伐珠单抗注射液1.25mg/眼组及IBI302各组增生纤维组织阳性强度无明显差异；就阳性分布而言，阳性对照组及IBI302各组较模型对照组减少，IBI302 1.25mg/眼组1只眼球未见阳性染色。
免疫组化结果显示：免疫组织化学染色结果表明IBI302各剂量组均能够减少脉络膜中CD31和C5b-9的表达，并且具有一定的剂量依赖性，贝伐珠单抗注射液对CD31的表达具有明显的抑制作用，对C5b-9的表达没有影响。具体结果见表24、表25、图8和图9。
表24.脉络膜免疫组化CD31检测结果
Figure PCTCN2020138598-appb-000032
表25.脉络膜免疫组化C5b-9检测结果
Figure PCTCN2020138598-appb-000033
Figure PCTCN2020138598-appb-000034
以上研究结果表明，在激光诱导恒河猴脉络膜血管新生模型中，IBI302能够明显减少光凝处荧光素渗漏、荧光斑面积和荧光斑数，降低荧光斑病损处视网膜厚度，使色素上皮恢复完整和连续，降低房水中VEGF水平，减轻纤维组织增生，抑制脉络膜中CD31和C5b-9的表达，IBI302比临床剂量贝伐珠单抗疗效更好。
安全药理学
50只恒河猴随机分配到5组，对照组、IBI302静脉注射8mg/猴组、IBI302玻璃体注射0.5mg/眼组、IBI302玻璃体注射2mg/眼组和IBI302玻璃体注射4mg/眼组。每组10只动物，雌雄各半。给药期及停药恢复期每天观察各组猴一般行为活动、精神状态、呼吸状态等；每周测定一次体温；第1次给药后约6、24、48、120小时，末次给药后约6小时，恢复期第4周及恢复期结束观察猴呼吸状态并检测II导联心电图及血压。
实验结果表明，给药期间及停药恢复期间，各组恒河猴一般状况良好，自主活动正常，也未见与供试品相关的行为学异常；研究期间各组恒河猴体温未见异常。首次给药后6、24、48、120小时，末次给药后约6小时，恢复期4周及恢复期结束，IBI302各组猴心率、QT及校正QT间期、PR间期、RR间期、P波时间、QRS波时间等心电图指标，收缩压、舒张压、平均压及肉眼观察呼吸状态均未见药物相关异常改变。提示恒河猴玻璃体注射IBI302 0.5mg/眼、2mg/眼或者4mg/眼或者静脉注射IBI302 8mg/猴对中枢神经系统、呼吸系统和心血管系统均无明显影响。
结论
药效学研究结论
IBI302一方面能够通过VID端特异性结合VEGF家族，抑制VEGF-A诱导的HUVEC的增殖；另一方面能够通过CID端特异性结合C3b和C4b，抑制补体旁路途径和经典途径介导的溶血，表明IBI302为双靶点特异性单克隆抗体，靶点明确，作用机制清楚。
IBI302通过双重阻断VEGF-A和补体介导的信号通路，抑制t-BHP刺激hRPE细胞释放VEGF-A、CCL2、C3a、C5a和MAC而发挥hRPE细胞保护作用；抑制激光诱导脉络膜血管新生小鼠表达VEGF、CCL2、TNF-α、C3a、C5a以及MAC，减轻脉络膜血管新生；抑制激光诱导脉络膜血管新生恒河猴表达VEGF、CD31和C5-b9，降低脉络膜血管新生和新生血管渗漏，降低视网膜厚度，保护色素上皮，表明IBI302药效确切。
药效学特征及意义
(1)双靶点：IBI302一方面通过VID端特异性结合VEGF-A165、VEGF-A121和PIGF，另一方面通过CID端特异性结合C3b和C4b，属于双靶向药物，靶点明确。
(2)作用机制清楚：IBI302的CID能够通过特异性结合C3b和C4b，抑制补体经典途 径和旁路途经的激活，减轻补体活化介导的炎症反应；VID端能够与VEGF家族结合阻断VEGF介导的信号通路，抑制血管内皮细胞的生存、增殖，从而抑制血管新生，降低血管渗透性，减少血管渗漏。
(3)疗效明确：在t-BHP诱导hRPE氧化应激模型显示IBI302对hRPE有显著地保护作用；在激光诱导小鼠和恒河猴脉络膜血管新生模型中，IBI302能产生明显抗血管新生和渗漏作用，提示IBI302具有非常好的临床疗效。
(4)比市售抗VEGF药物疗效更好：在t-BHP诱导hRPE氧化应激模型、激光诱导小鼠和恒河猴脉络膜血管新生模型中，IBI302均比阿柏西普或贝伐珠单抗疗效更好，提示IBI302可能比市售单独抗VEGF药物疗效更好。
(5)临床适应症：综合IBI302截至目前的临床前药理活性和类似药物的临床应用，IBI302在临床上拟用于湿性年龄相关性黄斑变性(AMD)的治疗。
实施例3：药代动力学研究
吸收
IBI302在恒河猴体内的药代动力学研究
在恒河猴单次静脉或者玻璃体注射IBI302的药代动力学研究中，27只恒河猴随机分为静脉注射组和玻璃体注射组，其中玻璃体注射组21只动物(雌性11只，雄性10只)，静脉注射组6只动物(雌性3只，雄性3只)。静脉注射组动物和玻璃体注射组的6只动物分别于给药前及给药后0.5h、1h、2h、4h、10h、24h、48h、72h、96h、144h、192h、240h、336h、504h采集血液，分离血清。玻璃体注射组于给药后分别取4h、24h、72h、168h、336h、504h的房水、玻璃体液、晶状体、角膜、虹膜、视网膜、脉络膜、巩膜、视神经及血清。详细实验设计见表26。
表26.实验设计表
Figure PCTCN2020138598-appb-000035
结果显示：玻璃体内注射IBI302后，药物半衰期T1/2显著大于静脉注射。眼部药物浓度显著高于血清药物浓度，系统毒性风险较小。从药物的眼部组织分布来看，药物主要分布在眼部组织的玻璃体、视网膜和脉络膜中，并且无论从末端消除半衰期T1/2还是平均驻留时间 MRT来看，药物在视网膜和脉络膜中均能停留较长的时间，从而发挥作用。
血清的药代动力学结果见表27和图10，眼部组织的药代动力学结果见表28、图11和12。
表27.恒河猴玻璃体内及静脉注射IBI302后血清的药代动力学参数
Figure PCTCN2020138598-appb-000036
N/A：不适用。
表28.恒河猴单次玻璃体内注射IBI302后眼部组织的药代动力学参数
Figure PCTCN2020138598-appb-000037
IBI302重复给药毒代动力学研究
在恒河猴玻璃体注射IBI302重复给药4周毒性研究中，50只恒河猴随机分配到5组，对照组、IBI302 8mg/只静脉注射组、IBI302 0.5mg/眼玻璃体注射组、IBI302 2mg/眼玻璃体注射组和IBI302 4mg/眼玻璃体注射组，每2周给药1次，连续给药3次，每组10只动物，雌雄各半。对照组分别于首次和末次给药前、给药后24h采集血液，制备血清。IBI302 8mg/只静脉注射组分别于首次和末次给药前、给药后5min、1h、4h、8h、24h、48h、72h、120h、168h和336h采集血液，制备血清。IBI302玻璃体注射给药组分别于首次和末次给药前、给药后1h、4h、8h、24h、48h、72h、120h和168h采集血液，制备血清。采用已建好的ELISA方法检测血清中IBI302浓度。具体实验设计见表29。
表29.实验设计概况
Figure PCTCN2020138598-appb-000038
Figure PCTCN2020138598-appb-000039
结果显示，恒河猴连续静脉注射IBI302后蓄积指数为0.7。恒河猴玻璃体内重复注射0.5mg/眼、2mg/眼、4mg/眼IBI302首次给药后，Cmax和AUC(0-t)倍比关系分别为1:4.4:5.5和1:6.3:13.4，有一定的剂量依赖关系；末次给药后，Cmax和AUC(0-t)倍比关系分别为1:2.3:7.9和1:15.7:54.6。IBI302各剂量蓄积系数分别为NA(末次个别动物未检测到药物，无法计算)、0.3和0.4。具体结果见表30和表31以及图13和图14。
表30.恒河猴首次静脉和玻璃体内注射IBI302后的毒代参数
Figure PCTCN2020138598-appb-000040
表31.恒河猴末次静脉注射及玻璃体内注射IBI302各剂量后的毒代参数
Figure PCTCN2020138598-appb-000041
以上结果表明，恒河猴玻璃体内重复注射IBI302，药物暴露量及达峰浓度随剂量增加而增加，呈现一定的剂量相关性。末次注射给药后血清中药物暴露量比首次给药后暴露量均有不同程度的降低，结合免疫原性结果表明部分动物产生了抗抗体，同时也表明重复给药后未见蓄积。
分布
见实施例1的IBI302在恒河猴体内的药代动力学研究。
代谢
根据ICH S6 Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals以及《治疗用生物制品非临床安全性技术审评一般原则》，IBI302为抗体药物，不需要开展代谢研究，因此并未开展IBI302代谢相关研究。
根据这类药物的代谢规律，预期IBI302的代谢产物为肽段或氨基酸。
排泄
未开展研究。
结论
药代动力学研究结论
恒河猴玻璃体注射IBI302后，IBI302在局部(眼部)范围内代谢速率显著低于全身，眼内局部药物浓度显著高于血清药物浓度，绝大部分药物均限制在眼部，系统毒性风险较小。药物的眼部暴露量表明玻璃体液、视网膜和脉络膜中药物分布最多，从而发挥药效作用。恒河猴玻璃体内重复注射IBI302，药物暴露量及达峰浓度随剂量增加而增加，呈现一定的剂量相关性，末次注射给药后血清中药物暴露量比首次给药后暴露量均有不同程度的降低，结合免疫原性结果表明部分动物产生了抗抗体，同时也表明重复给药后未见蓄积。
药代动力学特征及意义
(1)组织分布：玻璃体内注射IBI302后，绝大部分药物均限制在眼部，眼内局部药物浓度显著高于血清药物浓度，提示IBI302主要在眼部发挥药理活性，系统毒性风险较小。同时玻璃体液、视网膜和脉络膜中药物分布最多，符合药物主要作用于视网膜和脉络膜的设计目标。
(2)药效维持时间长：玻璃体注射IBI302后，无论从末端消除半衰期T1/2还是平均驻留时间MRT来看，药物在视网膜和脉络膜中均能停留较长的时间，从而有利于发挥作用。
(3)蓄积性：恒河猴静脉或者玻璃体重复注射IBI302，末次给药后的药物暴露量均低于首次给药，无蓄积性。
(4)免疫原性：恒河猴静脉或者玻璃体重复注射IBI302后，末次注射给药后血清中药物暴露量比首次给药后暴露量均有不同程度的降低，提示部分动物体内产生了针对IBI302的抗体。
实施例4：毒理学研究
单次给药毒性
恒河猴单次经玻璃体注射给予2mg/眼或4mg/眼剂量的IBI302，观察14天，所有猴未见明显毒性反应，表明恒河猴单次经玻璃体注射给予2mg/眼或4mg/眼剂量的IBI302能良好耐受。
重复给药毒性
50只恒河猴随机分配到5组，对照组、IBI302静脉注射8mg/猴组、IBI302玻璃体注射0.5mg/眼组、IBI302玻璃体注射2mg/眼组和IBI302玻璃体注射4mg/眼组。每组10只动物，雌雄各半。对照组和IBI302 4mg/眼组分别按100μl/眼的体积经双眼玻璃体注射给予0.9％氯化钠注射液和40mg/ml的IBI302，IBI302 0.5mg/眼组、2mg/眼组分别按50μl/眼的体积经双眼玻璃体注射给予10mg/ml和40mg/ml的IBI302。每2周给药1次，连续给药3次，停药恢复期为74天。具体实验设计见表32。
表32.恒河猴玻璃体重复注射IBI302毒性研究实验设计表
Figure PCTCN2020138598-appb-000042
Figure PCTCN2020138598-appb-000043
结果显示：
静脉组及各玻璃体注射组恒河猴体重、体温、摄食量、血液学、血生化、尿液指标、骨髓指标、心电血压、脏器重量及脏器系数以及除眼球、视神经以外的各脏器组织病理学检查等均未见异常改变。
玻璃体注射各剂量IBI302均可致恒河猴不同程度的眼内炎症，主要表现为：
0.5mg/眼组：给药期间，1/10恒河猴(右眼第一次给药后3天开始；左眼第2次给药后7天开始)，间接检眼镜检查发现玻璃体重度混浊/屈光介质混浊、眼底看不见或视网膜分支静脉白鞘样改变、出血灶、玻璃体混浊；裂隙灯检查发现前房渗出、眼压降低、玻璃体混浊、虹膜纹理不清、房水闪辉、细胞、角膜水肿混浊或前房渗出、AR、玻璃体混浊；眼压明显降低；白细胞计数(WBC)、单核细胞(MONO)、中性粒细胞及百分比(NEU、NEU％)可见明显升高；病理检查发现炎症细胞浸润、玻璃体变性。
恢复期第4周发现1/4视网膜分支血管周围白鞘样改变，至恢复期第6周基本恢复，病理检查发现仅个别眼见轻微炎症细胞浸润。
2mg/眼组：给药期间，1/10猴(左眼第2次给药后3天开始；右眼第3次给药后3天)间接检眼镜检查发现出现前房渗出、玻璃体重度混浊、眼底看不见或视网膜静脉血管周围白鞘样改变伴血管周围出血；裂隙灯检查检查发现前房渗出、瞳孔难以散大、球结膜水肿、混合充血、角膜混浊、房水闪辉、虹膜纹理不清、水肿或虹膜纹理不清、水肿；眼压明显降低；WBC、MONO、NEU、NEU％可见明显升高；大体解剖发现双侧眼球前房瞳孔区可见灰白色改变。另一只猴第3次给药后1天，玻璃体内出现血块(可能与注射相关)，至第3次给药后第6天可见玻璃体混浊，眼底看不见。眼电生理(ERG)检查发现给药结束雌猴明适应3.0(Cone-R)b波幅度降低，病理检查发现出现眼球单核细胞浸润、中性粒细胞浸润、玻璃体变性、视网膜/虹膜水肿及轻微视神经单核细胞浸润。
恢复期第4周2/4猴视网膜分支血管周围白鞘样改变，至恢复期第6周基本恢复；恢复期4、7周雌猴明适应3.0(Cone-R)b波幅度降低；病理检查仅个别眼出现轻微视神经、睫状体或视网膜单核细胞浸润。
4mg/眼组：给药期间，1/10猴(双眼，第3次给药后6天)间接眼底镜发现双眼玻璃体混浊；裂隙灯检查发现前房渗出，玻璃体混浊。另一只猴1只猴第1次给药后1天左眼出现前房渗出。给药结束雌猴明适应3.0(Cone-R)b波、暗适应3.0(Max-R)a波幅度降低。病理检查发现眼球单核细胞浸润、玻璃体变性、视网膜水肿及轻微视神经单核细胞浸润。
恢复期第4周，间接眼底镜发现3/4猴视网膜分支血管周围白鞘样改变，至恢复期第10周基本恢复；裂隙灯检查发现1只猴(1/4比例)双眼第5周出现玻璃体混浊，恢复期第6周恢复正常。恢复期4、7周雌猴明适应3.0(Cone-R)b波幅度降低，恢复期10周未见；恢复期第4周2/4只猴双眼OCT扫描可见眼底血管周围视网膜明显水肿、增厚，视网膜血管处反射增强，其下对应组织扫描成黑色阴影。恢复期7周未见。病理检查发现，仅个别眼出现轻 微视神经、睫状体或视网膜单核细胞浸润。
综上所述：恒河猴每2周1次、连续3次经玻璃体注射给予0.5mg/眼、2mg/眼或者4mg/眼剂量或静脉注射8mg/只剂量的IBI302，恢复74天。各剂量未见系统毒性，仅发现个别动物眼内炎症，恢复期结束，各组眼科检查眼内炎症基本恢复，眼组织病理学改变可见明显恢复。
遗传毒性
据根据ICH S6 Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals以及《治疗用生物制品非临床安全性技术审评一般原则》，IBI302为抗体药物，代谢产物为肽段或氨基酸，均不会对DNA和其他遗传物质产生影响，不需要开展遗传毒性研究，因此并未开展IBI302遗传毒性相关研究。
致癌性
本品采用玻璃体内注射给药，系统暴露量少，针对50岁以上的适应症人群，因此未开展致癌性研究。
生殖毒性
本品采用玻璃体内注射给药，系统暴露量少，针对50岁以上的适应症人群，生殖毒性安全风险担忧不大，因此未开展生殖毒性研究。
局部耐受性
日本大耳白兔每周1次连续4周滴眼给予0.5mg/眼或2mg/眼(浓度分别为10mg/ml或40mg/ml)剂量的IBI302，对兔眼无刺激性。
其他毒理试验
免疫原性
50只恒河猴随机分配到5组，对照组、IBI302静脉注射8mg/猴组、IBI302玻璃体注射0.5mg/眼组、IBI302玻璃体注射2mg/眼组和IBI302玻璃体注射4mg/眼组。每组10只动物，雌雄各半。每2周给药1次，连续给药3次，停药恢复期为74天。
免疫原性研究结果表明末次给药后，玻璃体注射2mg/眼、4mg/眼组雌、雄猴循环免疫复合物(CIC)升高，恢复期第4周，玻璃体注射4mg/眼组雌、雄猴CIC也可见升高趋势。各剂量组均有部分动物产生抗药抗体，经生物活性方法分析结果显示部分抗药抗体可能为中和抗体，表明IBI302具有一定的免疫原性。
免疫毒性
50只恒河猴随机分配到5组，对照组、IBI302静脉注射8mg/猴组、IBI302玻璃体注射0.5mg/眼组、IBI302玻璃体注射2mg/眼组和IBI302玻璃体注射4mg/眼组。每组10只动物，雌雄各半。每2周给药1次，连续给药3次，停药恢复期为74天。
在免疫毒性研究中，实验结果表明IBI302各组雌、雄猴其余免疫指标免疫球蛋白IgG、IgA、IgM和T淋巴细胞亚群CD3+、CD4+、CD8+均未见异常改变。给药结束及恢复期结束，IBI302各组猴脾脏和胸腺的绝对重量和脏体系数均未见异常改变，病理学检查亦未见异常改变。表明IBI302没有免疫毒性。
其他试验
组织交叉反应
用常规免疫组织化学方法(一步法)考察IBI302与正常人体和食蟹猴组织的免疫交叉反 应。实验结果表明：(1)在2μg/ml和20μg/ml浓度条件下，生物素化IBI302(Biotin-IBI302)和生物素化阿柏西普(Biotin-ZALTRAP)在重组人VEGF-A165蛋白涂片中可见特异性着色；Biotin-IBI302在人补体C4b蛋白涂片中可见特异性着色，Biotin-ZALTRAP在人补体C4b蛋白涂片中未见特异性着色；在阴性对照组中，未见Biotin-IBI302或Biotin-ZALTRAP特异性着色，且在试剂对照组(IBI302 placebo)中的所有蛋白涂片中未观察到着色，表明实验系统成立。(3)在2μg/ml和20μg/ml浓度条件下，IBI302和阿柏西普在正常人体和食蟹猴组织中的免疫组织化学染色未见特异性着色。
结论
毒理学研究结论
恒河猴单次经玻璃体注射给予2mg/眼或4mg/眼剂量的IBI302，未见明显毒性反应，耐受性良好。恒河猴每2周1次、连续3次经玻璃体注射给予0.5mg/眼、2mg/眼或者4mg/眼剂量或静脉注射8mg/只剂量的IBI302，恢复74天。结果显示未见明显系统毒性，玻璃体注射各剂量IBI302均可致猴不同程度的眼内炎症，并可见相应的眼和视神经炎症性病理改变。恢复期结束，各组眼科检查眼内炎症基本恢复，眼组织病理学改变可见明显恢复。静脉注射和玻璃体注射各剂量组均可产生抗药抗体，部分抗药抗体为中和抗体，恢复10周无法完全消除。眼内炎症可能是因为抗药抗体产生和免疫复合物形成相关，临床研究出现可能较低，风险较小。同时，日本大耳白兔每周1次连续4周滴眼给予0.5mg/眼或2mg/眼剂量的IBI302，对兔眼无刺激性。IBI302与正常人体组织和食蟹猴组织均无明显的组织交叉反应。
毒理学研究特征及意义
(1)单次给药：恒河猴单次经玻璃体注射给予2mg/眼或4mg/眼剂量的IBI302，未见明显毒性反应，同时对中枢神经系统、呼吸系统和心血管系统也均无明显影响。提示玻璃体单次注射给予IBI302具有很高的安全性。
(2)多次给药：恒河猴每2周1次、连续3次经玻璃体注射给予0.5mg/眼、2mg/眼或者4mg/眼剂量或静脉注射8mg/只剂量的IBI302，恢复74天。未见明显系统毒性。主要症状为玻璃体注射组可见不同程度的眼内炎症，并可见相应的眼和视神经炎症性病理改变，恢复期结束，各组眼科检查眼内炎症基本恢复，眼组织病理学改变可见明显恢复。提示IBI302的毒性反应仅发生于眼睛局部，同时具有可恢复性，具有很好的安全性。
(3)免疫毒性和免疫原性：恒河猴玻璃体重复注射IBI302未见任何免疫毒性。各剂量IBI302组均可产生抗药抗体，发生率较高。提示IBI302在恒河猴中具有一定的免疫原性，但仍建议在临床研究中密切关注。
(4)眼内炎：IBI302玻璃体注射出现的眼内炎症，与抗药抗体产生和免疫复合物形成有关，考虑到IBI302为全人源单克隆抗体，临床出现抗药抗体和免疫复合物可能性较低，因此眼内炎风险较小，但仍提醒临床研究中密切关注。
(5)局部刺激实验：日本大耳白兔每周1次连续4周滴眼给予0.5mg/眼或2mg/眼剂量的IBI302，对兔眼无刺激性。提示IBI302对眼睛无刺激性。
(6)组织特异性：IBI302与正常人体组织和食蟹猴组织均无明显的交叉反应，提示IBI302与正常人体组织无明显相互作用，靶点相关毒性非常低。
实施例5：在湿性年龄相关黄斑变性(AMD)患者中单次给药耐受性和安全性的剂量递 增I期临床研究
研究目的
评价湿性AMD患者单次玻璃体内注射IBI302的安全性、耐受性、生物活性及疗效。
药物
IBI302，规格0.2ml：2mg；0.2ml：8mg。
0.5mg/眼、2mg/眼和4mg/眼的剂量可从现有规格的研究药物中直接抽取。
其他剂量按以下步骤配制：
用注射器从药瓶中抽取固定体积的研究药物；
注入对应体积的溶媒瓶中；
拔出注射器，将溶媒与研究药物混匀；
用新的注射器从混匀的溶媒瓶中抽取50μl进行眼内注射。
各剂量组需要抽取的药物体积和对应的溶媒体积如下：
剂量组 药物规格 抽取药物体积 溶媒体积 抽取体积(眼内注射)
0.05mg/眼 0.2ml：2mg 90μl 760μl 50μl
0.15mg/眼 0.2ml：8mg 60μl 760μl 50μl
0.5mg/眼 0.2ml：2mg 50μl - 50μl
1mg/眼 0.2ml：8mg 760μl 760μl 50μl
2mg/眼 0.2ml：8mg 50μl - 50μl
4mg/眼 0.2ml：8mg 100μl - 100μl
研究设计
本研究为多中心、开放、剂量递增研究设计，继发于湿性AMD的活动性黄斑中心凹下脉络膜新生血管生成(CNV)患者可以入组本临床研究。每例受试者不可双眼均作为研究眼。合格受试者于研究第一天(D1)接受研究药物玻璃体内注射。给药后受试者应在研究中心接受安全性监测直至D2，确认无任何具有临床意义的异常情况，受试者才能离开研究中心。在D3-D6、D8、D15、D29、D43，受试者应回医院进行随访。参见图15的剂量递增示意图。
本研究共设6个剂量组：0.05mg/眼、0.15mg/眼、0.5mg/眼、1mg/眼、2mg/眼、4mg/眼。采用3+3剂量递增设计。以给药后14天内(即，D1给药后至D15检查完成)观察到剂量限制性毒性(DLT)作为剂量递增依据。将在0.15mg/眼至4mg/眼剂量组开展PK、PD和免疫原性研究，其中0.5mg/眼至4mg/眼剂量组，在完成剂量递增研究后需在最高耐受剂量(MTD)之前指定剂量组增加入组至6例受试者。参见图16的研究流程示意图。
剂量限制性毒性定义为DLT观察期内发生的以下任何一项与IBI302相关的不良事件(AE)：
轻度及以上眼内炎症(参见附录2)；
给药后眼压持续升高≥10mmHg超过60min；
轻度及以上与眼内炎症不相关的急性视力下降(参见附录2)；
中度及以上玻璃体内出血(参见附录2)；
颅内出血或其他有临床意义的眼外其他部位出血；
研究者认为与研究药物有关的严重不良事件(SAE)。
附录2：眼部预期不良事件分级判断标准
Figure PCTCN2020138598-appb-000044
附录3：前房闪辉/细胞数分级标准
参考Hogan等人(Hogan MJ,Kimura SJ,Thygeson P.)Am J.Ophthalmol 1959；47:155–70的 前部葡萄膜炎和葡萄膜炎的体征和症状分级标准。该文献的全部内容整体引入本文作为参考。
附录4：玻璃体内细胞数和出血程度分级标准
Figure PCTCN2020138598-appb-000045
研究人群
50岁及以上男性或女性；
研究眼继发于湿性AMD的活动性黄斑中心凹下CNV，病损大小≤12个视盘直径；
糖尿病视网膜病变早期治疗研究组(ETDRS)视力表测定研究眼最佳矫正视力(BCVA)在10-73个字母范围内；
经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm；
眼睛中部和晶状体不浑浊，可获得清晰的立体眼底摄影；
育龄期女性受试者或伴侣为育龄期女性的男性受试者，同意从筛选期开始直至治疗结束后3个月采取有效的避孕措施；
自愿签署书面知情同意书，而且能够遵守方案规定的访视及相关程序。
给药方案
受试者按入组顺序依次分别接受0.05mg/眼、0.15mg/眼、0.5mg/眼、1mg/眼、2mg/眼或4mg/眼剂量的IBI302单次眼内注射。
评价指标
安全性评价指标：
BCVA的下降情况；
给药后眼压较基线的变化；
眼内炎症的发生率和严重程度；
治疗后，生命体征、体格检查结果和实验室结果较基线的变化；
所有不良事件(AE)、治疗期不良事件(TEAE)和严重不良事件(SAE)的发生率、与研究药物的相关性以及严重程度等。
有效性评价指标：
D8、D15、D29和D43时，眼科OCT检查中心视网膜厚度较基线变化；
D43时荧光素血管造影(FA)检查CNV特点及CNV面积较基线变化；
D2、D4、D6、D8、D15、D29和D43时BCVA较基线的变化。
免疫原性评价指标：
抗药抗体(ADA)和中和抗体(NAb)阳性率。
PK评价指标：
PK参数包括但不限于：0到时间t的药时曲线下面积(AUC0-t)、0到无穷大的药时曲线下面积(AUC0-∞)、峰浓度(Cmax)、达峰时间(Tmax)、清除率(CL)、表观分布容积(V)、半衰期(t1/2)、平均滞留时间(MRT)等。
PD评价指标：
游离VEGF浓度、总VEGF浓度、补体相关指标(C3、C4、C5及其裂解片段)以及炎症相关指标(IL-1b、IL-6)。
统计学方法
样本量：
本研究共设6个剂量组，采用3+3剂量递增设计对这6个剂量水平进行评估，在0.5mg/眼、1mg/眼、2mg/眼和4mg/眼剂量组，完成剂量递增研究后需在MTD之前指定剂量组增加入组至6例受试者，预计总共需要入组约30～36例湿性AMD患者。
统计分析方法：
以描述性统计为主，原则上不进行组间比较。按剂量组总结在湿性AMD患者玻璃体内单次注射IBI302的安全性指标、PK/PD参数、免疫原性和疗效指标。
连续性变量采用例数、均数、标准差、中位数、最小值和最大值进行描述；分类变量采用频数、百分比进行描述。此外，对于疗效性指标，还将提供不同剂量组下指标均数的95％置信区间。
实验结果
IBI302治疗nAMD的初步有效性数据来自多中心、开放、剂量递增研究设计的I期单药剂量递增研究CIBI302A101，共设6个剂量组：0.05mg/眼、0.15mg/眼、0.5mg/眼、1mg/眼、2mg/眼、4mg/眼，合格受试者于研究第一天(D1)接受研究药物玻璃体内注射。采用3+3剂量递增设计，其中0.5mg/眼至4mg/眼剂量组，在完成剂量递增研究后需在最高耐受剂量之前指定剂量组增加入组至6例受试者。共入组31例nAMD受试者，并接受研究药物 给药。
1.安全性和耐受性
无以下实验室检查异常：血常规、血生化、肝功能、尿常规
无生命体征以及体格检查的异常表现，仅1例受试者(1/15)出现给药后BCVA↓>5字母，且于给药后第六天恢复至基线水平无SAE发生，包括眼内炎、动脉栓塞事件、视网膜脱离、视网膜色素上皮撕裂最常见的AE是球结膜下出血，(6/15,40％，自限性，不影响视力)，有两例受试者出现给药后眼内压IOP>21mmHg(正常值上限)，但为自限性，24小时内恢复正常。
2.最佳矫正视力(BCVA)
使用ETDRS视力表在4米的测试距离下开始检查受试者坐位时的视力。观察正确识别字母数，结果显示正确识别字母数呈上升趋势，初期试验结果见图17。其中，BCVA的数值为字母正确识别数；ΔBCVA％＝AVERVGE(本次随访正确识别字母数-第一次随访正确识别字母数)×100％。该图中显示各剂量组的平均值。
在整个研究的第2、4、6、8、15、29、43天，即给药后第1、3、5、7、14、28、42天，所有受试者研究眼平均最佳矫正视力(BCVA)较基线分别提高2.1±3.87、2.8±4.49、3.8±4.69、5.5±6.19、5.9±6.86、6.0±8.15、6.1±7.47个字母。
研究眼的BCVA在D2、4、6、8、15、29、43天，分别有83.9％、74.2％、87.1％、83.9％、87.1％、77.4％、71.0％的受试者较基线有提高；分别有19.4％、29.0％、32.3％、48.4％、54.8％、48.4％、48.4％的受试者提高≥5个字母；分别有6.5％、3.2％、16.1％、22.6％、25.8％、25.8％、22.6％的受试者提高≥10个字母，提高≥15个字母的比例分别有0、3.2％、3.2％、9.7％、12.9％、19.4％、12.9％。各剂量组受试者在各访视时的研究眼BCVA检查结果均值较基线均有提高。检查结果随时间的变化趋势、检查结果较基线的改变值随时间的变化趋势分别见图19、图20。图中显示各剂量组的平均值。
3.光学相干断层扫描(OCT)
出院后随访D8、D15、D29和D43，进行OCT检查。给药后所有受试者均出现了网膜内积液(IRF)、网膜下积液(SRF)不同程度的吸收，且吸收程度与快慢呈一定的剂量依赖关系。
4.中心视网膜厚度(CST)
出院后随访D8、D15、D29和D43，检测中心视网膜厚度。给药后所有受试者均出现了中心视网膜厚度降低，且厚度降低趋势呈一定的剂量依赖关系。中心视网膜厚度(CST)变化趋势及与基线比较的初期试验结果见图18。该图中显示各剂量组的平均值。
在整个研究的D8、15、29、43天，即给药后第7、14、28、42天，所有受试者研究眼OCT检查CST分别平均减少了100±80.62、122.8±100.48、141.2±153.66、66.1±205.05μm。
给药后各剂量组研究眼OCT检查CST均有减少，厚度减少的程度在给药后第28天达到最大。从0.5mg剂量组开始，厚度的减少幅度显著高于0.05mg与0.15mg剂量组。4mg剂量组在D29时，减少幅度最大，达到328.5μm。各剂量组研究眼CST检查结果随时间变化趋 势见图21，各剂量组研究眼CST检查结果较基线的变化随时间变化趋势见图22。图中显示各剂量组的平均值。
具体而言，在黄斑区视网膜厚度改善上，使用了OCT测量的中央区1mm的视网膜平均厚度(CST)作为观察指标。所有剂量组在给药后第7天，即出现CST的下降。在给药后第7天(即D8)，各剂量组(0.05mg、0.15mg、0.5mg、1mg、2mg和4mg)分别较基线下降了21.3±29.69μm、8.3±90.47μm、136.8±89.87μm、142±100.97μm、90.6±41.94μm和117.5±44.70μm，较基线下降百分比分别为5％、2％、22％、29％、17％、23％。其中，0.5mg及以上剂量组，CST改善的程度显著高于0.05mg和0.15mg剂量组。在给药后第28天(即D29)，各剂量组(0.05mg、0.15mg、0.5mg、1mg、2mg和4mg)分别较基线下降了8.0±67.64μm、23.7±79.53μm、171.8±131.88μm、203.7±159.66μm、115.3±154.06μm和328.5±191.63μm，较基线下降百分比分别为1.7％、4.4％、27.4％、41.1％、22.0％、63.3％。其中，0.5mg及以上剂量组，CST改善的程度显著高于0.05mg和0.15mg剂量组。CST改善的效应持续至研究结束。
5.FA检查脉络膜新血管形成(CNV)面积
在整个研究中，给药后第42天，所有受试者的研究眼FA检查CNV面积平均减少0.524±3.7806mm 2。除2mg剂量组外，各剂量组在给药后42天时的CNV面积均减少。对侧眼的CNV面积平均减少0.352±1.7891mm 2。其中，在给药后第42天，0.05mg剂量组相对于基线下降0.980±0.4857mm 2，0.15mg剂量组相对于基线下降4.963±9.9122mm 2，0.5mg剂量组相对于基线下降0.365±1.7140mm 2，1mg剂量组相对于基线下降0.820±1.7530mm 2，4mg剂量组相对于基线下降0.327±0.4206mm 2。上述数据采用首次给予研究药物前(包括筛选期的数据)的最后一次非缺失值作为基线。变化值＝测量值-基线值。

Claims (26)

  1. 在个体中预防或治疗年龄相关性黄斑变性(AMD)的方法，该方法包括向所述个体施用抑制VEGF途径和补体途径的融合蛋白。
  2. 根据权利要求1所述的方法，所述融合蛋白包含SEQ ID NO：1的序列。
  3. 根据权利要求1所述的方法，所述融合蛋白的序列为SEQ ID NO：1所示的序列。
  4. 根据权利要求1-3任一项所述的方法，其中所述的AMD为湿性AMD。
  5. 根据权利要求4所述的方法，所述的湿性AMD具有以下的一种或多种症状和体征：视力下降、视物变形、中心暗点、阅读困难、黄斑区网膜肿胀、眼底出血、新生血管形成、瘢痕纤维化形成、地图样萎缩，或其任意组合。
  6. 根据权利要求4所述的方法，所述的湿性AMD具有脉络膜新血管形成(CNV)。
  7. 根据权利要求4所述的方法，所述的湿性AMD为具有以下特征的AMD：
    (1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
    (2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
  8. 根据上述权利要求任一项所述的方法，其中所述个体为人类个体，其具有上述权利要求任一项所述的疾病、症状和/或特征。
  9. 根据上述权利要求任一项所述的方法，其中所述个体为人类个体，其为湿性AMD患者并伴有地图样萎缩。
  10. 根据上述权利要求任一项所述的方法，其中所述个体患有具备以下特征的湿性AMD：
    (1)研究眼继发于湿性AMD的活动性黄斑中心凹下CNV；和/或
    (2)经光学相干断层成像(OCT)扫描，中心视网膜厚度≥250μm。
  11. 根据上述权利要求任一项所述的方法，其中所述治疗可在个体中实现以下一种或多种效果：
    1)中心视网膜厚度较基线降低；
    2)脉络膜新生血管生成(CNV)面积降低；
    3)最佳矫正视力(BCVA)较基线提高；
    4)湿性AMD患者地图样萎缩发生时间滞后；和/或
    5)湿性AMD患者地图样萎缩发生时间减慢。
  12. 根据上述权利要求任一项所述的方法，其中所述治疗可在个体中实现以下一种或多种效果：
    1)提高最佳矫正视力(BCVA)；和/或
    2)降低中心视网膜厚度；和/或
    3)降低脉络膜新生血管生成(CNV)面积。
  13. 根据上述权利要求任一项所述的方法，其中所述治疗可在个体中实现以下一种或多种效果：
    1)最佳矫正视力(BCVA)与基线水平相比提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母；和/或
    2)最佳矫正视力(BCVA)与基线水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％，和/或
    3)中心视网膜厚度(CST)较基线水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者较基线水平降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％；和/或
    4)脉络膜新生血管生成(CNV)面积与基线水平相比降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与基线水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
  14. 根据上述权利要求任一项所述的方法，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的中心视网膜厚度水平的给药前水平相比，给药后六周或八周，该个体的中心视网膜厚度水平降低25μm以上，例如降低25-350μm，优选50-300μm，更优选50-200μm，例如降低25、50、75、100、125、150、175、200、225、250、275、300、325、350μm，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％， 更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
  15. 根据上述权利要求任一项所述的方法，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的脉络膜新生血管生成(CNV)面积的给药前水平相比，给药后六周或八周，该个体的脉络膜新生血管生成(CNV)面积降低0.1mm 2以上，例如降低0.1-20mm 2，优选0.2-5mm 2，更优选0.3-2mm 2，例如降低0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5、10、10.5、11、11.5、12、12.5、13、13.5、14、14.5、15、15.5、16、16.5、17、17.5、18、18.5、19、19.5或20mm 2，或者与给药前水平相比降低5％以上，例如降低10％-70％，优选10％-60％，更优选30％-50％，更优选40-50％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％或70％。
  16. 根据上述权利要求任一项所述的方法，所述方法包括向所述个体以约0.01-10mg/眼，优选0.05-8mg/眼，更优选0.05-6mg/眼，更优选0.5-5mg/眼，更优选1-5mg/眼，更优选2-5mg/眼的单次剂量，每天一次、每两天一次、每周两次、每周一次、每两周一次、每四周一次、每五周一次或每六周一次玻璃体内或静脉内施用所述融合蛋白，与个体的最佳矫正视力(BCVA)的给药前水平相比，给药后六周或八周，该个体的最佳矫正视力(BCVA)提高至少1个ETDRS字母，优选提高至少5个ETDRS字母，更优选提高5-35个ETDRS字母，例如提高5-25个ETDRS字母，例如提高5-10个、5-15个、5-20个、10-15个、10-20个、10-25个、15-20个或15-25个字母，例如提高5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34或35个字母，或者与给药前的水平相比提高5％以上，优选提高10％以上，例如提高10％-200％，优选10％-150％，更优选10％-100％，更优选20％-100％、更优选50％-100％、60％-100％、20％-80％或30％-70％，更优选50％-70％，更优选60％-70％，例如5％、10％、15％、20％、25％、30％、35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％、100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％。
  17. 抑制VEGF途径和补体途径的融合蛋白在制备用于在个体中预防或治疗年龄相关性黄斑变性(AMD)的药物中的用途，其中所述融合蛋白包含SEQ ID NO：1的序列。
  18. 根据权利要求17所述的用途，其中所述融合蛋白的序列为SEQ ID NO：1所示的序列。
  19. 根据权利要求17-18任一项所述的用途，其中所述AMD如权利要求4-7任一项所述。
  20. 根据权利要求17-19任一项所述的用途，其中所述个体如权利要求8-10任一项所述。
  21. 一种单次药物剂量单元，其特征在于包含抑制VEGF途径和补体途径的融合蛋白，其中所述融合蛋白包含SEQ ID NO：1的序列。
  22. 根据权利要求21所述的单次药物剂量单元，其中所述融合蛋白的序列为SEQ ID NO：1所示的序列。
  23. 根据权利要求21或22所述的单次药物剂量单元，其中所述单次药物剂量单元包含以下剂量的所述融合蛋白：0.01-10mg，优选0.05-8mg，更优选0.05-6mg，更优选0.5-5mg，更优选1-5mg，更优选2-5mg，例如0.5-2mg、1-3mg、2-4mg、2-3mg或3-5mg，例如0.05mg、0.1mg、0.15mg、0.2mg、0.25mg、0.3mg、0.35mg、0.4mg、0.45mg、0.5mg、0.55mg、0.6mg、0.65mg、0.7mg、0.75mg、0.8mg、0.85mg、0.9mg、0.95mg、1mg、1.1mg、1.2mg、1.3mg、1.4mg、1.5mg、1.6mg、1.7mg、1.8mg、1.9mg、2mg、2.1mg、2.2mg、2.3mg、2.4mg、2.5mg、2.6mg、2.7mg、2.8mg、2.9mg、3mg、3.1mg、3.2mg、3.3mg、3.4mg、3.5mg、3.6mg、3.7mg、3.8mg、3.9mg、4mg、4.1mg、4.2mg、4.3mg、4.4mg、4.5mg、4.6mg、4.7mg、4.8mg、4.9mg、5mg、5.1mg、5.2mg、5.3mg、5.4mg、5.5mg、5.6mg、5.7mg、5.8mg、5.9mg、6mg、6.1mg、6.2mg、6.3mg、6.4mg、6.5mg、6.6mg、6.7mg、6.8mg、6.9mg、7mg、7.1mg、7.2mg、7.3mg、7.4mg、7.5mg、7.6mg、7.7mg、7.8mg、7.9mg、8mg、8.5mg、9mg、9.5mg、10mg，优选2mg或4mg。
  24. 一种成套药物包装盒，其特征在于，包含权利要求21-23中任一项所述的融合蛋白。
  25. 根据权利要求21-23任一项所述的单次药物剂量单元或权利要求24所述的成套药盒在制备预防或治疗年龄相关性黄斑变性(AMD)的药物中的用途。
  26. 根据权利要求21-23任一项所述的单次药物剂量单元或权利要求24所述的成套药盒，其用于预防或治疗年龄相关性黄斑变性(AMD)。
PCT/CN2020/138598 2019-12-24 2020-12-23 融合蛋白在治疗年龄相关性黄斑变性中的应用 WO2021129658A1 (zh)

Priority Applications (6)

Application Number Priority Date Filing Date Title
CA3165887A CA3165887A1 (en) 2019-12-24 2020-12-23 Use of fusion protein in treatment of age-related macular degeneration
CN202080090126.6A CN114867489A (zh) 2019-12-24 2020-12-23 融合蛋白在治疗年龄相关性黄斑变性中的应用
EP20904512.9A EP4082559A4 (en) 2019-12-24 2020-12-23 APPLICATION OF FUSION PROTEIN IN THE TREATMENT OF AGE-RELATED MACULAR DEGENERATION
JP2022538928A JP2023509880A (ja) 2019-12-24 2020-12-23 加齢黄斑変性の治療における融合タンパク質の使用
AU2020414152A AU2020414152A1 (en) 2019-12-24 2020-12-23 Application of fusion protein in treating age-related macular degeneration
US17/788,181 US20230056821A1 (en) 2019-12-24 2020-12-23 Use of fusion protein in treatment of age-related macular degeneration

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN201911346311.2 2019-12-24
CN201911346311 2019-12-24
CN202011441025.7 2020-12-11
CN202011441025 2020-12-11

Publications (1)

Publication Number Publication Date
WO2021129658A1 true WO2021129658A1 (zh) 2021-07-01

Family

ID=76575187

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/138598 WO2021129658A1 (zh) 2019-12-24 2020-12-23 融合蛋白在治疗年龄相关性黄斑变性中的应用

Country Status (8)

Country Link
US (1) US20230056821A1 (zh)
EP (1) EP4082559A4 (zh)
JP (1) JP2023509880A (zh)
CN (1) CN114867489A (zh)
AU (1) AU2020414152A1 (zh)
CA (1) CA3165887A1 (zh)
TW (1) TW202128746A (zh)
WO (1) WO2021129658A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117467025B (zh) * 2023-12-28 2024-04-16 上海鼎新基因科技有限公司 一种抗vegf和补体双功能融合蛋白及其应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013082563A1 (en) 2011-12-01 2013-06-06 Protevobio, Inc. Protein inhibitors to complement and vegf pathways and methods of use thereof
CN104940926A (zh) * 2014-09-25 2015-09-30 信达生物制药(苏州)有限公司 重组融合蛋白制剂
CN105435222A (zh) * 2014-09-25 2016-03-30 信达生物制药(苏州)有限公司 重组融合蛋白制剂

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013082563A1 (en) 2011-12-01 2013-06-06 Protevobio, Inc. Protein inhibitors to complement and vegf pathways and methods of use thereof
CN104159926A (zh) * 2011-12-01 2014-11-19 普腾生技有限公司 补体和vegf途径的蛋白质抑制剂及其使用方法
CN104940926A (zh) * 2014-09-25 2015-09-30 信达生物制药(苏州)有限公司 重组融合蛋白制剂
CN105435222A (zh) * 2014-09-25 2016-03-30 信达生物制药(苏州)有限公司 重组融合蛋白制剂

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
HOGAN ET AL., AM J. OPHTHALMOL, vol. 47, 1959, pages 155 - 70
INNOVENT BIOLOGICS SUZHOU CO., LTD.: "A Dose Escalation Study of IBI302 in Patients with Wet Age-related Macular Degeneration", no. NCT03814291, 24 January 2019 (2019-01-24), pages 1 - 4, XP009537740, Retrieved from the Internet <URL:https://clinicaltrials.gov/ct2/show/NCT03814291> *
REN XINYI; LI JIA; XU XIANXING; WANG CHUNMING; CHENG YUANGUO: "IBI302, a Promising Candidate for AMD Treatment, Targeting Both the VEGF and Complement System with High Binding Affinity in Vitro and Effective Targeting of the Ocular Tissue in Healthy Rhesus Monkeys", EXPERIMENTAL EYE RESEARCH, vol. 145, 23 February 2016 (2016-02-23), pages 352 - 358, XP029516892, ISSN: 0014-4835, DOI: 10.1016/j.exer.2016.02.004 *
See also references of EP4082559A4
YULIANG FENG ; MING ZHANG ; CHUNMING WANG ; JIA LI ; QIAORONG DAN: "Inhibitory Effects of IBI302 on Experimental Choroidal Neovascularization", CHINESE JOURNAL OF OCULAR FUNDUS DISEASES, vol. 32, no. 2, 31 March 2016 (2016-03-31), pages 177 - 183, XP055823769, ISSN: 1005-1015, DOI: 10.3760/cma.j.issn.1005-1015.2016.02.015 *

Also Published As

Publication number Publication date
AU2020414152A1 (en) 2022-08-04
EP4082559A4 (en) 2024-01-03
US20230056821A1 (en) 2023-02-23
CN114867489A (zh) 2022-08-05
JP2023509880A (ja) 2023-03-10
EP4082559A1 (en) 2022-11-02
CA3165887A1 (en) 2021-07-01
TW202128746A (zh) 2021-08-01

Similar Documents

Publication Publication Date Title
De Cogan et al. Topical delivery of anti-VEGF drugs to the ocular posterior segment using cell-penetrating peptides
US20210205410A1 (en) Use of a vegf antagonist to treat angiogenic eye disorders
Zhang et al. Recombinant anti-vascular endothelial growth factor fusion protein efficiently suppresses choridal neovasularization in monkeys
US10208124B2 (en) Anti-CD160 specific antibodies for the treatment of eye disorders based on neoangiogenesis
CN109715189A (zh) 靶向VE-PTP(HPTP-β)的人源化单克隆抗体
US20120301478A1 (en) Pharmaceutical for Preventing or Treating Disorders Accompanied by Ocular Angiogenesis and/or Elevated Ocular Vascular Permeability
EP4185318A1 (en) Extended, high dose vegf antagonist regimens for treatment of angiogenic eye disorders
WO2021129658A1 (zh) 融合蛋白在治疗年龄相关性黄斑变性中的应用
US20120101033A1 (en) Retinitis pigmentosa treatment
US11478465B2 (en) Compounds for treatment of eye diseases associated with excessive vascularisation
EP4028128A1 (en) Methods for treating ocular diseases
Ulutas et al. Does intravitreal injection of aflibercept affect the corneal endothelium?
KR20220051791A (ko) Fas 신호전달 억제용 펩타이드를 포함하는 황반변성 예방 또는 치료용 조성물
Ghoz Ocular neovascularization: pathological changes in cornea, conjunctiva and retina
Hakan et al. Evaluation Of The Effectiveness Of Intravitreal Anti-VEGF Injection With Intravitreal Dexamethasone Implant In Patients With Treatment-Resistant Diabetic Macular Edema
EA046420B1 (ru) Применение антагониста vegf для лечения ангиогенных глазных заболеваний
Fernandez-Lloris Association for Research in Vision and Ophthalmology (ARVO)-2015 Annual Meeting. Denver, Colorado, USA-May 3-7, 2015
Ferrer et al. Ophthalmic therapeutics: new basic research and clinical studies
Al-Shamahi Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO). Seattle, Washington, USA-May 5-9, 2013

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20904512

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2022538928

Country of ref document: JP

Kind code of ref document: A

Ref document number: 3165887

Country of ref document: CA

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2020904512

Country of ref document: EP

Effective date: 20220725

ENP Entry into the national phase

Ref document number: 2020414152

Country of ref document: AU

Date of ref document: 20201223

Kind code of ref document: A