Complement factor I dosing regimen for treatment of ocular diseases

Abstract

Provided herein are dosing regimens for treating ocular conditions, such as age-related macular degeneration (AMD). These regimens include administering to a subject in need thereof a fusion construct comprising complement factor I (CFI) and a binding partner at specified doses and frequencies.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 338,392, filed on May 4, 2022, the contents of which are incorporated herein by reference in their entirety.

[0003] References to Electronic Sequence Listings

[0004] The contents of the electronic sequence listing (VTEX_708_01WO_SeqList_ST26.xml; size: 18,753 bytes; and creation date: May 3, 2023) are incorporated herein by reference in their entirety. Background Art

[0005] The complement system includes the classical pathway, the lectin pathway and the alternative pathway, and is tightly controlled by a variety of regulatory factors. Complement factor I (CFI) is a type of such regulatory factor, and is used to regulate the complement system by cutting C4b protein and C3b protein, thereby inactivating these proteins. Such cutting results in the inhibition of the classical pathway, the lectin pathway and the alternative pathway, respectively, thereby ultimately preventing the assembly of C3 convertase and C5 convertase. CFI is encoded as a zymogen, which is then activated as a heterodimeric glycoprotein by proteolytic cleavage, and the heterodimeric glycoprotein has a heavy chain (A chain) and a light chain (B chain) connected by a disulfide bond. CFI is processed into its active form by furin after translation to produce a double-chain mature protein.

[0006] CFI dysregulation, CFI mutation and dysfunction (i.e., genetic variants) or CFI deficiency are associated with diseases involving the complement system. Dysregulation or overactivation of the complement system is associated with eye diseases such as age-related macular degeneration (AMD). The progression of AMD leads to macular damage, retinal pigment epithelium (RPE) atrophy and photoreceptor loss, resulting in blurred vision and eventual loss of vision. Rare genetic variants carrying CFI and / or low CFI levels are associated with AMD and the progression of the disease. According to the National Eye Institute of the National Institutes of Health (NIH), AMD is the leading cause of vision loss in adults, with more than 190 million people affected worldwide in 2020. There are two types of AMD: wet AMD and dry AMD. Wet AMD is less common and is characterized by abnormal growth of blood vessels at the back of the eye, thereby damaging the macula. Dry AMD is characterized by thinning of the macula and can progress to map atrophy (GA), which is characterized by progressive and irreversible loss of retinal tissue. GA is the leading cause of blindness and affects more than 5 million people worldwide. There is a need for methods to modulate or inhibit the complement system to maintain regulatory balance for the treatment of ocular diseases such as AMD and GA. Provided herein are compositions and methods for addressing dysfunction and / or dysregulation in the complement system. Summary of the invention

[0007] In one aspect, the present invention provides a method for treating or preventing an ocular condition in a human subject in need thereof, the method comprising administering to the subject a fusion construct comprising a complement factor I or a variant thereof comprising the amino acid sequence of SEQ ID NO.5 and a binding partner, wherein the route of administration is intraocular, and wherein the fusion construct is administered at a dose of about 0.1 mg per eye to about 10 mg per eye and / or at a frequency of about every 1 month to 4 months. In some embodiments, the binding partner is serum albumin or a variant thereof comprising the amino acid sequence of SEQ ID NO.7. In some embodiments, the CFI variant has at least 70%, at least 80%, at least 90%, or at least 95% identity to the amino acid sequence of SEQ ID NO.5. In some embodiments, CFI variants include CFI of SEQ ID NO:5 comprising one or more of substitutions T377G, N422K, E457G, and N531G; CFI of SEQ ID NO:5 comprising substitutions N422K, E457G, and N531G; or CFI of SEQ ID NO:5 comprising substitutions T377G, E457G, and N531G. In some embodiments, the binding partner is a cofactor of CFI, such as factor H (FH) or complement receptor 1 (CR1). In some embodiments, the factor H binding partner comprises the amino acid sequence of SEQ ID NO.22 or SEQ ID NO:23, a domain thereof, or a portion of its domain. In some embodiments, the binding partner comprises CR1, a domain thereof, or a portion of its domain. The CFI-FH fusion protein may also comprise serum albumin, such as serum albumin comprising the amino acid sequence of SEQ ID NO.7, or a variant thereof. The CFI-CR1 fusion protein may further comprise serum albumin, such as serum albumin comprising the amino acid sequence of SEQ ID NO.7 or a variant thereof. In some embodiments, the route of administration is intravitreal. In some embodiments, the ocular condition is dry age-related macular degeneration (AMD). In some embodiments, the ocular condition is wet age-related macular degeneration (AMD). In some embodiments, the ocular condition is geographic atrophy (GA). In some embodiments, the subject exhibits GA. In some embodiments, the human subject carries one or more mutations of the CFI gene selected from the group consisting of: G119R, L131R, V152M, G162D, R187Y, R187T, T203I, A240G, A258T, G287R, A300T, R317W, R339Q, V412M, P553S, K441R, R339Ter, R317Q, G261D,In some embodiments, the ocular disorder is caused by one or more mutations in the CFI gene of the subject, the mutations selected from the group consisting of Phe13Val, Val20Ile, His40Arg, Cys43Phe, Asp44Asn, Pro50Ala, Cys54Arg, Cys54*, Ile55Phe, Pro64Leu, Asn70Thr, Thr72Ser, Phe82Ser, Pro83Gln, Ser90Asn, Cys106Arg, Thr107Ala, Glu109Ala, His118Arg, Gly119Arg, Gly125Arg, Va l127Ala, Val129Gly, Leu131Arg, Met138Val, Met138Ile, Trp145*, Asn151Ser, Val152Met, Gly162Asp, Asp164Val, Asn177Ile, His183Arg, Val184M et, Arg187*, Arg187Gln, Gly188Ala, Phe198Leu, Arg202Ile, Thr203Ile, Tyr206Asn, Gln217His, Ser221Tyr, Asp224Asn, Cys229Arg, Val230Met, Va l230Glu, Ala240Gly, Cys247Gly, Gly248Glu, Asp249Glu, Ala258Thr, Gly261Ser, Gly261Asp, Gly263Val, Lys267Asn, Gly280Asp, Gly287Arg, Glu29 0Asp、Ala300Thr、Glu303Lys、Glu305*、Ile306Val、Ile306Ser、Asp310Glu、Arg317Trp、Arg317Gln、Cys327Arg、Gly328Arg、Arg339*、Arg339Gln、Il e340Thr, Gly342Glu, Arg345Gln, Gly349Arg, Val355Met, Ala356Pro, Ile357Met, Gly362Ala, Tyr369Ser, Trp374Cys, Arg389His, Trp399Arg, Pro40 2Ser, Asp403Asn, Arg406Cys, Arg406His, Tyr411*, Val412Met, Ile416Leu, His418Leu, Gly424Asp, Asp429Glu, Ala431Thr, Ile433Val, Ile433Thr,Lys441Arg, Arg448Cys, Ala452Ser, Trp456Leu, Tyr459Ser, Gln462His, Cys467Arg, Arg474*, Arg474Gln, Asp477His, Gly487Cys, Ile492Leu, Gly500Arg, Arg502Cys, Gly512Ser, Asp519Asn, Lys522Thr, Asp524Val, Met532Val, Asn536Lys, Trp541*, Gly542Ser, Val543Ala, Trp546*, Glu548Gln, Pro553Ser, Glu554Val, Ser570Thr, Ile578Thr and Gln580*. In some embodiments, the dose is about 0.1 mg per eye to about 10 mg per eye. In some embodiments, the dose is about 0.1 mg per eye, 0.5 mg per eye, 1 mg per eye, 5 mg per eye, or 10 mg per eye. In some embodiments, the frequency is about every 1 month. In some embodiments, the frequency is about every 2 months, 3 months, or 4 months. In some embodiments, the fusion construct is administered to one eye. In some embodiments, the fusion construct is administered to both eyes sequentially. In some embodiments, the fusion construct is administered to both eyes simultaneously. In some embodiments, the intraocular pressure of the subject is stable after treatment. In some embodiments, the retinal thickness of the subject is stable after treatment. In some embodiments, the activity exhibited by the fusion construct in the subject is at least equivalent to the activity exhibited by the plasma-derived or naturally occurring CFI protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 is a diagram depicting a model of an exemplary CFI-albumin (eg, serum albumin, such as human serum albumin (HSA)) fusion construct comprising serum albumin fused to CFI, wherein the CFI comprises wild-type CFI.

[0009] Figure 2A is a graph showing the amount of C3a detected in samples from vitreous humor following intravitreal injection (IVT) of a fusion construct comprising wild-type CFI and human serum albumin (CFI-HSA) at a dose of 250 μg or 500 μg in an African green monkey (AGM) primate model.

[0010] Figure 2B is a graph showing the C3b / iC3b ratio detected in samples from vitreous humor following intravitreal injection (IVT) of a 1.0 mg dose of a fusion construct comprising wild-type CFI and human serum albumin (CFI-HSA) in the AGM primate model.

[0011] Figure 3A is a graph depicting vitreous pharmacokinetics (PK) in AGM. Constructs containing CFI-HSA were injected intravitreally at 0.25 mg per eye or 1 mg per eye. The amount of injected construct was measured over time. Figure 3B is a graph depicting vitreous PK modeling in humans based on PK measured for AGM.

[0012] Figure 4A is a graph depicting clinical scores obtained by slit-lamp for intravitreal injections of CFI-HSA in nonhuman primates. Figure 4B is a graph depicting intraocular pressure measured over time in nonhuman primates following intravitreal injection of CFI-HSA. Figure 4C is a graph depicting retinal thickness measured over time in nonhuman primates following intravitreal injection of CFI-HSA. DETAILED DESCRIPTION

[0013] The present disclosure provides dosage regimens of fusion constructs comprising CFI and a binding partner (including, for example, HSA, factor H or CR1) that can be used to modulate the signaling and amplification of the complement system in the ocular system. The present disclosure also provides methods of preparing and using these fusion constructs, for example, for treating ocular diseases or conditions associated with complement dysregulation, such as dry age-related macular degeneration (AMD), wet AMD, and geographic atrophy (GA).

[0014] I. Complement Factor I Fusion Constructs

[0015] Provided herein are complement factor I (CFI) fusion constructs, such fusion constructs comprising:

[0016] (a) wild-type CFI or a variant thereof, and

[0017] (b) Binding partner.

[0018] In some embodiments, the binding partner comprises serum albumin or a variant thereof. In some embodiments, the binding partner comprises a cofactor of CFI, such as Factor H or CR1.

[0019] In some embodiments, the fusion construct comprises:

[0020] (a) complement factor I comprising the amino acid sequence of SEQ ID NO.5 or a variant thereof, and

[0021] (b) serum albumin comprising the amino acid sequence of SEQ ID NO. 7 or a variant thereof.

[0022] In some embodiments, the fusion construct comprises:

[0023] (a) a complement factor I comprising the amino acid sequence of SEQ ID NO.5 or a variant thereof,

[0024] (b) a factor H cofactor or a domain thereof comprising the amino acid sequence of SEQ ID NO. 22 or 23, and

[0025] (c) optionally, serum albumin comprising the amino acid sequence of SEQ ID NO. 7 or a variant thereof.

[0026] In some embodiments, the fusion construct comprises:

[0027] (a) a complement factor I comprising the amino acid sequence of SEQ ID NO.5 or a variant thereof,

[0028] (b) a CR1 cofactor or a domain thereof, and

[0029] (c) optionally, serum albumin comprising the amino acid sequence of SEQ ID NO. 7 or a variant thereof.

[0030] As used herein, wild-type CFI refers to any naturally occurring full-length or nearly full-length CFI that is non-pathogenic, with or without a signal sequence, and can be from any species.

[0031] In some embodiments, the wild-type CFI is plasma derived. In some embodiments, the wild-type CFI is human wild-type CFI. In some embodiments, the wild-type human CFI with a signal sequence comprises the amino acid sequence set forth in SEQ ID NO: 1 (as shown in Table 1 below). In some embodiments, the wild-type CFI is human CFI. In some embodiments, the wild-type human CFI does not comprise a signal sequence. In some embodiments, the wild-type CFI without a signal sequence comprises the amino acid sequence set forth in SEQ ID NO: 5 (as shown in Table 1 below).

[0032] Wild-type CFI comprises a heavy chain and a light chain, which are also referred to as the A-chain and the B-chain, respectively. Figure 1 A schematic diagram showing the CFI of these two chains is depicted.

[0033] Exemplary base molecules used as components of CFI fusion constructs are provided in Table 1. The base molecules of Table 1 are used to generate the CFI fusion constructs disclosed herein. The base molecules of Table 1 can be another part of a fusion construct, further described below.

[0034] Provided herein are fusion constructs comprising variants of CFI, the variants comprising one or more modifications relative to wild-type CFI, referred to herein as "CFI variants." As used herein, "modifications" relative to wild-type CFI include: deletion of one or more amino acid residues, deletion of one or more domains, substitution of one or more amino acid residues, insertion (i.e., addition) of one or more amino acid residues, insertion (i.e., addition) of one or more domains, inversion of one or more domains, and substitution of one or more domains.

[0035] Variants of CFI may include those that confer additional CFI activity to one or more of the classical pathway, the lectin pathway, and the alternative pathway. In some embodiments, modifications relative to wild-type CFI include any one or more of the following: deletion of one or more amino acid residues, deletion of one or more CFI domains, substitution of one or more amino acid residues, insertion of one or more amino acid residues, insertion of one or more CFI domains, and exchange of one or more CFI domains. PCT / US2021 / 037278 and PCT / US2022 / 082179 provide variants of CFI and are incorporated herein by reference in their entirety.

[0036] In some embodiments, the CFI variant comprises any one or more domains of CFI selected from the following: serine protease domain (SPD), factor I membrane attack complex (FIMAC) domain, SRCR domain, low-density lipoprotein receptor 1 (LDLr1) domain, and low-density lipoprotein receptor 2 (LDLr2) domain.

[0037] In some embodiments, the CFI variant comprises at least one modification corresponding to wild-type human CFI. In some embodiments, the CFI variant comprises at least one modification corresponding to wild-type non-human CFI. In some embodiments, the CFI variant comprises at least one modification corresponding to wild-type CFI having an amino acid sequence as set forth in SEQ ID NO: 1 or SEQ ID NO: 5.

[0038] In some embodiments, the CFI variant is a chimera comprising one or more domains from human CFI, and wherein the human CFI further comprises one or more amino acid residues substituted for amino acid residues from the corresponding region of the non-human species CFI. In some embodiments, the non-human species is a mouse. In some embodiments, the CFI variant is a chimera, and wherein the modification comprises substitution of one or more amino acid residues of CFI with amino acid residues from the corresponding region of a non-CFI serine protease. In some embodiments, the non-CFI serine protease is trypsin.

[0039] In some embodiments, the CFI variant comprises an A chain and a B chain, wherein the CFI variant comprises one or more modifications at an interface of the A chain and the B chain.

[0040] In some embodiments, the CFI variant comprises a modification at any one or more positions corresponding to positions K14, Y20, D26, F29, R35, E38, M220, K221, S250, L304, P305, K306, L307, and S308 in the CFI having the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI variant comprises one or more substitutions selected from the group consisting of K14A, Y20A, Y20F, D26A, F29A, R35A, E38A, M220A, K221Q, S250A, S250L, L304G, P305G, K306G, L307G, and S308G, wherein the positions correspond to positions in the CFI having the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI variant comprises one or more of a combination of substitutions M220A; K221Q and L304G; P305G; K306G; L307G; S308G; wherein the positions correspond to positions in the CFI having the amino acid sequence shown in SEQ ID NO:5.

[0041] In some embodiments, the CFI variant comprises one or more modifications in the C-terminal region of the CFI variant. In some embodiments, the CFI variant comprises a modification at any one or more positions corresponding to positions T377, W381, P384, Y403, A405, G406, Y408, Q409, D425, G556, R557, P558, P559, I560, and Y563 in the CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI variant comprises one or more substitutions selected from the group consisting of T377G, W381A, P384A, P384G, Y403F, A405S, G406R, G406A, Y408L, Q409D, Q409H, D425A, D425K, D425R, G556A, G556S, R557A, R557K, P558G, P558L, P558S, F559L, I560V, and Y563H and / or a deletion of P384, wherein the positions correspond to positions in the CFI having the amino acid sequence shown in SEQ ID NO:5.

[0042] In some embodiments, the CFI variant comprises one or more modifications at one or more N-linked glycosylation sites of CFI. In some embodiments, the CFI variant comprises a modification at any one or more positions corresponding to positions N52, N85, N159, N446, N476, and N518 in the CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI variant comprises one or more substitutions selected from the group consisting of: N52Q, N85Q, N159Q, N446Q, N476Q, and N518Q, wherein the positions correspond to the positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI variant comprises one or more of a combination of substitutions selected from the group consisting of: N52Q; N85Q; N159Q, N446Q; N476Q; N518Q; wherein the positions correspond to the positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0043] In some embodiments, the CFI variant comprises one or more modifications in the SPD domain of CFI. In some embodiments, the CFI variant comprises one or more modifications in the SPD domain of CFI. In some embodiments, the CFI variant comprises one or more modifications in the SPD domain of CFI at positions K14, K312, R314, I322, V323, K326, R327, A328, K340, D341, G344, I345, T346, A361, L364, Y372, W381, P384, V390, N402, N404, G406, Y408, Q409, E416, K418, N422, D425 , E457, K458, R456, E461, R462, F464, S465, Q467, W468, G469, T495, Y496, D497, S499, I500, A502, K504, D506, S507, E530, N531, E530, N531, G533, K534, P535, E536, and F537 comprise modifications.

[0044] In some embodiments, the CFI variant comprises a substitution of the autolytic loop of CFI for the autolytic loop of trypsin, wherein the autolytic loop occurs between positions corresponding to position 456 and position 465 in CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0045] In some embodiments, the CFI variant comprises one or more substitutions selected from the group consisting of K14A, K312A, R314A, I322T, I322Y, I322V, V323I, V323G, V323A, K326A, R327A, R327P, R327N, A328C, K340G, D341A, G344R, G344K, G344Y, I345G, T346R, T346K, T346H , A361G, L364G, Y372G, W381K, W381G, P384A, P384G, V390G, N402E, N404G, G406D, G406E, G406F, G406 H, G406I, G406K, G406L, G406M, G406N, G406P, G406Q, G406S, G406T, G406V, G406W, G406Y, Y408L, Y40 8F, Y408G, Y408P, Y408D, Y408A, Y408N, Y408T, Y408K, Y408R, Y408H, Y408I, Y408E, Y408M, Y408Q, Y4 08S, Y408W, Y408V, Q409G, E416A, K418G, N422K, D425A, D425K, D425R, D425G, R456A, R456N, E457G, E 457A, E457D, E457F, E457H, E457I, E457K, E457L, E457M, E457N, E457P, E457Q, E457R, E457S, E457T, E457W, E457Y, E457V, K458A, E461Q, E461K, E461R, E461H, E461G, E461A, E461D, E461F, E461I, E461LE461M, E461N, E461P, E461S, E461T, E461W, E461Y, E461V, R462K, R462A, R46 2D, F464Y, S465G, Q467K, Q467R, W468C, G469L, T495F, Y496L, D497E, S499G, I500K, A502S, K504Q, K504E, K504R, K504A, K504G, K504L, K504P, K504H, K50 4D, K504F, K504I, K504M, K504N, K504S, K504T, K504V, K504W, K504Y, D506A, D506V, D506E, D506G, S507A, E530D, E530G, E530F, E530Y, N531G, N531A, E530D, E530G, E530F, E530Y, E530R, E530K, N531D, N531E, N531F, N531H, N531I, N531K, N531L, N531M, N531P, N531Q, N531R, N531S, N531T, N531V, N531W, N531Y, G533A, K534Q, P535A, P535K, E536N, E536A, F537K, and F537R, wherein the positions are identical to those having SEQ The position in the CFI of the amino acid sequence shown in ID NO:5 corresponds to that in the CFI.

[0046] In some embodiments, the CFI variant comprises one or more of the following combination substitutions: K326A; R327A, N531G; P535A, E457G; E461Q; R462K; F464Y, Y408L; N531G; E457G, Y408L; N531G; E457G; E461Q, Y408L; N531G; E457G; E461Q-R462K; F464Y, Y408L; N531G; P535A, K14A; D425R, E530D; N531G; G533A; K534Q; P535K; E536N, A502S; K504Q; F537K, T495F; Y4 96L; D497E; S499G; I500K, G533A; K534Q; P535K; E536N; F537K, T495F; Y496 L; D497E; S499G; I500K; G533A; K534Q; P535K; E536N; F537K, Q467K; F537K, E530G; N531G, E530D; F537K, E457G; E461Q, E457G; E461G, Y408L; N531G; E4 57G; E461Q, N531G; E457G; E461Q, I322V; V323I, I322V; V323I; R327P, A328C ;W468C, A328C; W468C; K326Y; R327N, Y408L; N531G; E461Q, Y408L; N531G; E 457G; E461Q; R462K, Y408L; N531G; E457G; E461Q; F464Y, Y408L; N531G; E45 7G; R462K; F464Y, Y408L; N531G; E461Q; R462K; F464Y, Y408L; E457G; E461Q ;R462K; F464Y, E457G; N531G; E461Q; R462K; F464Y, Y408L; E457G; E461Q; R4 62K, N531G; E457G; E461Q; F464Y, E416A; D425R, Y408L; N531G; E457G; E461 Q; R462K; F464Y; S507A, E457G; E461G, K312A; R314A, G469L; R456N; E457T; K 458A, G469L; R456N; K458A, G469L; R456N; K458A; E461G, G469L; R456N; K45 8A; E461G; F537K, G406D; Y408L, G406D; N531G, G406D; P535A, G406D; Y408L;N531G, G406D; Y408L; P535A, G406D; N531G; P535A, G406D; Y408L; N531G; P53 5A, K340G; I345G, L364G; Y372G, W381G; V390G, W381G; P384A; V390G, W381G; P384G; V390G, N404G; Q409G, K418G; D425G, T346R; K504E; E530R, T346K; K50 4D; E530K, G344R; Y408L; N531G, G344K; Y408L; N531G, T346R; Y408L; N531G, T346K; Y408L; N531G, K504D; Y408L; N531G, K504E; Y408L; N531G, Y408L; E530R; N531G, Y408L; E530K; N531G, T346R; Y408L; K504E; E530R; N531G, T346K; Y408L; K504D; E530K; N531G, Y408L; S507A; N531G, Y408L; N531G; E457G; E461Q; R462K; F464Y; S507A, E457G; S507A and N531G; P535A; S507A; wherein the position is consistent with the The position in the CFI of the amino acid sequence shown in ID NO:5 corresponds to that in the CFI. ;

[0047] In some embodiments, the CFI variant comprises one or more modifications at the active site of CFI. In some embodiments, the CFI variant comprises a modification at a position corresponding to position S507 in CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI variant comprises substitution S507A, wherein the position corresponds to position S507 in CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0048] In some embodiments, the CFI variant comprises an A chain and a B chain, wherein the CFI variant comprises a structural arrangement of (A chain)-(optional linker)-(B chain) from the N-terminus to the C-terminus. In some embodiments, the CFI variant comprises an A chain and a B chain, wherein the CFI variant comprises a structural arrangement of (B chain)-(optional linker)-(A chain) from the N-terminus to the C-terminus. In some embodiments, the CFI variant comprises a modification at one or more of C309 and C435, wherein the positions correspond to positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0049] In some embodiments, the CFI variant comprises the substitutions C309S; C435S, wherein the positions correspond to positions in the CFI having the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the B chain and the A chain are further linked by a disulfide bond.

[0050] In some embodiments, the CFI variant is more activatable than wild-type CFI. In some embodiments, the CFI variant comprises a modification at any one or more positions corresponding to positions I317, R318, R319, K320, and R321 in the CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI variant comprises one or more substitutions selected from the group consisting of I317D, R318D, R319D, K320D, and R321K, wherein the positions correspond to positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0051] In some embodiments, the CFI variant comprises substitutions I317D, R318D, R319D, K320D, and R321K, wherein the positions correspond to positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0052] In some embodiments, the CFI variant comprises one or more of the combinations of substitutions selected from: Y408; N531G, E38A; D425R, Y20F; D425R, S250A; D425R, Y408F; N531G, Y408L; N531G; E457G; E461Q; R462K; F464Y, K14A; Y20F, K14A; E38A, K14A; S250A, K14A; D425A, Y20F; E38A, Y20F; S250A, Y20F; D425A, E38A; S250A, E38A; D425A, S250A; D425A, K14A; N531G; P535A, Y20F; N531G; P535A, E38A; N531G; P535A, S250A; N531G; P535A, D425A; N531G; P535A, Y20F; Y408L; N531G; E457G; E461Q; R462K; F464Y, E38A; Y408L; N531G; E457G; E461Q; R462K; F464Y, S250A; Y408L; N531G; E457G; E461Q; R462K; F464Y, D425R; Y408L; N531G; E457G; E461Q; R462K; F464Y, Y20F; E38A; S250A; D425A, Y20F; E38A; S250A; D425A; Y408L; N531G; E457G; E461Q; R462K; F464Y, Y20F; E38A; S250A; D425A; Y408L; N531G; E457G; E461Q, I317D; R318D; R319D; K320D; R321K; E457G; E461Q-R462K; F464Y, I317D; R318D; R319D; K320D; R321K; E457G; E461Q-R462K; F464Y, I317D; R318D; R319D; K320D; R321K; Y408L; N531G; E457G; E461Q; R462K; F464Y, K504D; Y408L; N531G, K504E; Y408L; N531G; E457G; N531G; D425K, Y408F; N531G, Y408L; E457G; N531G; D425K, Y408L; E457G; P535G; D425K, Y408L; E457G; N531G; K534Q, Y408L; N531G, R462K; F464Y and Y408L; P535G;D425K, wherein the position corresponds to a position in CFI having an amino acid sequence as shown in SEQ ID NO:5. ;

[0053] In some embodiments, the CFI variant is at positions K14, K312, R314, I322, V323, K326, R327, A328, T377, K340, D341, G344, I345, T346, A361, L364, Y372, W381, P384, V390, N402, N404, G406, Y408, Q409, E416, K418, N422, D425 , E457, K458, R456, E461, R462, F464, S465, Q467, W468, G469, T495, Y496, D497, S499, I500, A502, K504, D506, S507, E530, N531, E530, N531, G533, K534, P535, E536, and F537.

[0054] In some embodiments, the CFI variant comprises or consists of any one or more substitutions selected from the group consisting of K14A, K312A, R314A, I322T, I322Y, I322V, V323I, V323G, V323A, K326A, R327A, R327P, R327N, A328C, T377G, K340G, D341A, G344R, G344K, G344Y, I345G, T346R, T346 K, T346H, A361G, L364G, Y372G, W381K, W381G, P384A, P384G, V390G, N402E, N404G, G406D, G406E, G406F , G406H, G406I, G406K, G406L, G406M, G406N, G406P, G406Q, G406S, G406T, G406V, G406W, G406Y, Y408L, Y408F, Y408G, Y408P, Y408D, Y408A, Y408N, Y408T, Y408K, Y408R, Y408H, Y408I, Y408E, Y408M, Y408Q, Y 408S, Y408W, Y408Y, Y408V, Q409G, E416A, K418G, N422K, D425A, D425K, D425R, D425G, R456A, R456N, E4 57G, E457A, E457D, E457F, E457H, E457I, E457K, E457L, E457M, E457N, E457P, E457Q, E457R, E457S, E45 7T, E457W, E457Y, E457V, K458A, E461Q, E461K, E461R, E461H, E461G, E461A, E461D, E461F, E461I, E461LE461M, E461N, E461P, E461S, E461T, E461W, E461Y, E461V, R462K, R462A, R46 2D, F464Y, S465G, Q467K, Q467R, W468C, G469L, T495F, Y496L, D497E, S499G, I500K, A502S, K504Q, K504E, K504R, K504A, K504G, K504L, K504P, K504H, K50 4D, K504F, K504I, K504M, K504N, K504S, K504T, K504V, K504W, K504Y, D506A, D 506V, D506E, D506G, S507A, E530D, E530G, E530F, E530Y, N531G, N531A, E530 D. E530G, E530F, E530Y, E530R, E530K, N531D, N531E, N531F, N531H, N531I, N 531K, N531L, N531M, N531P, N531Q, N531R, N531S, N531T, N531V, N531W, N531Y, G533A, K534Q, P535A, P535K, E536N, E536A, F537K and F537R, wherein the positions correspond to the positions in the CFI having the amino acid sequence shown in SEQ ID NO:5.

[0055] In some embodiments, the CFI variant comprises or consists of any one or more of the following combination substitutions selected from: N422K+E457G+N531G, T377G+E457G+N531G, K326A+327A R327A, E457G+N531G, N531G+P535A, E457G+E461Q+R462K+F464Y, Y408L+N531G+E457G, Y408L+N531G+E457G+E461Q, Y408L+N531G+E45 7G+E461Q-R462K+F464Y, Y408L+N531G+P535A, K14A+D425R, E530D+N531G+G533A+K534Q+P535K+E536N, A502S+K504Q+F537K, T495F+Y4 96L+D497E+S499G+I500K, G533A+K534Q+P535K+E536N+F537K, T495F+Y496L+D497E+S499G+I500K+G533A+K534Q+P535K+E536N+F537K , Q467K+F537K, E530G+N531G, E530D+F537K, E457G+E461Q, E457G+E461G, Y408L+N531G+E457G+E461Q, N531G+E457G+E461Q, I322V+V32 3I, I322V+V323I+R327P, A328C+W468C, A328C+W468C+K326Y+R327N, Y408L+N531G+E461Q, Y408L+N531G+E457G+E461Q+R462K, Y408L+ N531G+E457G+E461Q+F464Y, Y408L+N531G+E457G+R462K+F464Y, Y408L+N531G+E461Q+R462K+F464Y, Y408L+E457G+E461Q+R462K+F464 Y、E457G+N531G+E461Q+R462K+F464Y、Y408L+E457G+E461Q+R462K、N531G+E457G+E461Q+F464Y、E416A+D425R、Y408L+N531G+E457G+E4 61Q+R462K+F464Y+S507A, E457G+E461G, K312A+R314A, G469L+R456N+E457T+K458A, G469L+R456N+K458A, G469L+R456N+K458A+E461G,G469L+R456N+K458A+E461G+F537K, G406D+Y408L, G406D+N531G, G406D+P535A, G406D+ Y408L+N531G, G406D+Y408L+P535A, G406D+N531G+P535A, G406D+Y408L+N531G+P535A, K 340G+I345G, L364G+Y372G, W381G+V390G, W381G+P384A+V390G, W381G+P384G+V390G, N4 04G+Q409G, K418G+D425G, T346R+K504E+E530R, T346K+K504D+E530K, G344R+Y408L+N53 1G, G344K+Y408L+N531G, T346R+Y408L+N531G, T346K+Y408L+N531G, K504D+Y408L+N53 1G, K504E+Y408L+N531G, Y408L+E530R+N531G, Y408L+E530K+N531G, T346R+Y408L+K504 E+E530R+N531G, T346K+Y408L+K504D+E530K+N531G, Y408L+S507A+N531G, Y408L+N531G+E457G+E461Q+R462K+F464Y+S507A, E457G+S507A, and N531G+P535A+S507A, wherein the positions correspond to the positions in the CFI having the amino acid sequence set forth in SEQ ID NO:5. For example, in some embodiments, the CFI variant comprises or consists of a substitution selected from the group consisting of: N422K+E457G+N531G. For example, in some embodiments, the CFI variant comprises or consists of a substitution selected from the group consisting of: T377G+E457G+N531G. ,

[0056] In some embodiments, the fusion construct comprises CFI and albumin, and the albumin is human serum albumin (HSA), and the CFI is wild-type CFI, and such fusion constructs are referred to herein as "CFI-HSA". Exemplary CFI-HSA of the present disclosure has the amino acid sequence of SEQ ID NO: 21, or a sequence having at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO. 21.

[0057] In some embodiments, the CFI of the fusion construct comprises the amino acid sequence of SEQ ID NO. 5 or a variant thereof. In some embodiments, the variant has at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO. 5.

[0058] In some embodiments, the serum albumin of the fusion construct comprises the amino acid sequence of SEQ ID NO. 7 or a variant thereof. In some embodiments, the variant has at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO. 7.

[0059] In some embodiments, the binding partner of CFI in the fusion construct includes at least one domain or a portion of a domain of complement receptor 1 (CR1). In some embodiments, the at least one CR1 domain includes CCP domains 15-17 of CR1. In some embodiments, the at least one CR1 domain includes CCP domains 1-3 of CR1. In some embodiments, the fusion construct of the present disclosure comprising at least one CR1 domain also includes a fusion with albumin. In some embodiments, the fusion construct of the present disclosure comprising at least one CR1 domain and / or at least one domain of factor H also includes a fusion with albumin. In some embodiments, the at least one CR1 domain includes CR1 CCP domain 15. In some embodiments, the at least one CR1 domain includes CR1 CCP domain 16. In some embodiments, the at least one CR1 domain includes CR1 CCP domain 17. In some embodiments, the at least one CR1 domain includes CR1 CCP domains 15-16. In some embodiments, the at least one CR1 domain includes CR1 CCP domains 16-17. In some embodiments, an exemplary fusion construct comprises CFI with modification N531G fused to CCP domains 15-17 of CR1. In some embodiments, an exemplary fusion construct comprises CFI fused to CCP domain of CR1 and further fused to albumin.

[0060] In some embodiments, the binding partner of CFI in the fusion construct includes at least one domain or a portion of a domain of factor H. In some embodiments, the at least one factor H domain includes any one or more of complement control protein (CCP) domains 1-20 of factor H. In some embodiments, the amino acid sequence of the at least one factor H domain is or is derived from the sequence shown in SEQ ID NO:22 (Table 1). In some embodiments, the at least one factor H domain includes each of CCP domains 1-20 of factor H. In some embodiments, the at least one factor H domain includes CCP1, CCP2, CCP3, and CCP4. In some embodiments, the at least one factor H domain includes CCP2, CCP3, and CCP4. In some embodiments, the at least one factor H domain includes CCP2 and CCP3. In some embodiments, the amino acid sequence of at least one domain of factor H is or is derived from the sequence shown in SEQ ID NO:23 (Table 1). In some embodiments, the at least one factor H domain includes CCP domains 1-4 and 19-20 of factor H. In some embodiments, an exemplary fusion construct comprises CFI fused to the CCP domain of FH, and further fused to albumin.

[0061] Exemplary CFI-HSA fusion constructs of the present disclosure include, as shown from N-terminus to C-terminus:

[0062] (a) N-terminal HSA-GGSSGG linker (SEQ ID NO:6) - CFI SEQ ID NO:5 comprising substitutions N422K, E457G and N531G.

[0063] (b) N-terminal HSA-GGSSGG linker (SEQ ID NO:6) - CFI SEQ ID NO:5 comprising substitutions T377G, E457G and N531G.

[0064] Exemplary CR1-containing fusion constructs of the present disclosure include HSA, CFI and human CR1 domains. Examples of such fusion constructs are:

[0065] (a) N-terminal HSA-GGSSGG linker (SEQ ID NO: 6) - CFI SEQ ID NO: 5 comprising substitutions N422K, E457G and N531G -

[0066] GGSSGGSSGGSSGGSSGGSSGG linker (SEQ ID NO:24) - human CR1 domains CCP15, CCP16 and CCP17.

[0067] The components of the fusion constructs of the present disclosure (CFI and binding partner) can be held together by an optional linker. They can have any suitable length of at least one amino acid. The linker can be a flexible linker and can be a peptide of about 1 to about 20 amino acid residues in length, wherein the amino acid residues may include glycine residues. The linker may also optionally contain a serine residue. Exemplary flexible linkers may include, but are not limited to, glycine polymers, glycine-serine polymers, glycine-alanine polymers, alanine-serine polymers, or any other suitable flexible linker known in the art. An exemplary linker is (GGSS) nGG (SEQ ID NO: 25), wherein n is any number from about 1 to about 25. In some embodiments, the linker is a protease-sensitive cleavable linker. Exemplary linkers for joining fusion constructs can be 1-50, 5-50, 10-50, 15-50, 20-22, 22-24, 20-50, 25-50, 1-20, 2-20, 3-20, 4-20, 5-20, 6-20, 7-20, 8-20, 9-20, 10-20, 3-15, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 4-15, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-15, 5-10, 5-9, 5-8, 5-7, 5-6, 6-15, 6-10, 6-9, 6-8, or 6-7 amino acids in length.

[0068] In some embodiments, CFI-HSA can have an extended half-life relative to CFI that is not part of a fusion construct. Exemplary CFI-HSA constructs can be generated by linking albumin to wild-type CFI with a flexible linker. In some embodiments, CFI-HSA comprises the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity thereto.

[0069] In some embodiments, the fusion construct comprises the structural arrangement from N-terminus to C-terminus of (albumin)-(optional linker)-(WT CFI).

[0070] In some embodiments, the fusion construct comprises the structural arrangement from N-terminus to C-terminus: (WT CFI)-(optional linker)-(albumin).

[0071] In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:7 and SEQ ID NO:5, or variants thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity to SEQ ID NO:5, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO:7)-(optional linker)-(SEQ ID NO:5) from N-terminus to C-terminus. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:5 and SEQ ID NO:7, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO:7)-(linker)-(SEQ ID NO:5) from N-terminus to C-terminus. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO:7)-(SEQ ID NO:6)-(SEQ ID NO:5) from N-terminus to C-terminus. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 7, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO: 5)-(optional linker)-(SEQ ID NO: 7) from the N-terminus to the C-terminus. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 7, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO: 5)-(linker)-(SEQ ID NO: 7) from the N-terminus to the C-terminus. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, wherein the fusion construct comprises a structural arrangement of (SEQ ID NO: 5)-(SEQ ID NO: 6)-(SEQ ID NO: 7) from the N-terminus to the C-terminus.

[0072] In some embodiments, the fusion construct comprises the amino acid sequence shown in SEQ ID NO: 21, or an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95% or at least 99% identity thereto. In some embodiments, the fusion construct consists of the amino acid sequence shown in SEQ ID NO: 21. In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 7. In some embodiments, it is noted that the fusion of albumin to wild-type CFI (e.g., N-terminal albumin fusion) provides solubility and promotes the activation of CFI-HSA. When CFI is activated as a mature two-chain protein with furin after translation and a comparison of activation is made between CFI-HSA and wild-type CFI (WT-CFI) without albumin, it is observed that furin activates CFI-HSA significantly better and almost completely. It was observed that the CFI-HSA protein remained as a monomer with no evidence of aggregates. Amino-terminal HSA fusion has significant and unexpected benefits for maintaining the solubility, monodispersity and efficient furin activation of the CFI-HSA construct. There is a significant improvement in bioavailability through, for example, improved half-life.

[0073] Thus, provided herein are methods of increasing CFI activation, the methods comprising fusing HSA to wild-type CFI, wherein the fusion is an N-terminal fusion prior to activation with furin; and activating with furin. In some embodiments, activation with furin is performed in cells during recombinant production of the CFI fusion constructs disclosed herein. In some embodiments, activation with furin is performed in vitro.

[0074] Turning to Table 1, SEQ ID NO: 1 is the amino acid sequence of wild-type plasma-derived human CFI (referred to as "CFI-PD"), and has a leader sequence. The wild-type CFI for fusion with the second component may comprise the amino acid sequence of SEQ ID NO: 5, which does not comprise the leader sequence present in SEQ ID NO: 1. The mouse Ig kappa chain V-III region MOPC 63 leader sequence (SEQ ID NO: 2) can instead be used to recombinantly produce any of the CFI fusion constructs provided herein. In some embodiments, provided herein is a CFI fusion construct comprising at least one CFI domain, wherein the at least one CFI domain comprises the amino acid sequence set forth in SEQ ID NO: 5.

[0075] Table 1: Components of exemplary CFI fusion constructs

[0076]

[0077]

[0078]

[0079] II. Generation of CFI Fusion Constructs

[0080] Cell lines can be developed to express the production of CFI fusion constructs described herein. Cell lines used to produce fusion constructs can be produced using any host cell capable of expressing the fusion constructs described herein. The host cell can be a mammalian cell, an insect cell, a fungal cell, a plant cell, and / or a bacterial cell. In order to express the CFI fusion construct, the host cell line can be transiently or stably transfected or transduced with an expression vector encoding the fusion construct. The vector can be, for example, a plasmid or a viral vector. In some embodiments, the host cell line is a mammalian cell line. In some embodiments, the host cell is a Chinese hamster ovary (CHO) cell.

[0081] The CFI fusion constructs described herein can be recombinantly expressed in mammalian cell lines known in the art for producing biologics, such as Chinese hamster ovary (CHO) cells. Mammalian cells can be transfected or transduced with expression vectors encoding the fusion constructs described herein using any method known in the art.

[0082] Provided herein are methods for producing and purifying the CFI fusion constructs described herein. The CFI fusion constructs described herein can be purified from conditioned medium by standard methods known in the art. In some embodiments, the CFI fusion constructs can be purified by chromatography on an affinity matrix. In some embodiments, the affinity matrix is ​​CaptureSelect TM Human albumin affinity matrix. In some embodiments, the CFI fusion construct can be purified by chromatography on a cation and / or anion exchange matrix and optionally size exclusion chromatography. The CFI fusion construct can be optimally buffer exchanged into any suitable buffer known in the art. Purity can be assessed by any method known in the art, including gel electrophoresis, orthogonal HPLC methods, staining, and spectrophotometric techniques.

[0083] III. CFI Fusion Constructs - Methods of Use

[0084] A. Application

[0085] The CFI fusion constructs of the present disclosure can be used to modulate the complement system in the eye.

[0086] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of modulating both the classical and lectin complement pathways in the eye.

[0087] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of modulating the alternative complement pathway in the eye.

[0088] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing the expansion of the complement system in the eye.

[0089] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the cleavage of C3b in the eye.

[0090] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the cleavage of C4b in the eye.

[0091] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the production of C4c in the eye.

[0092] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the production of iC3b in the eye.

[0093] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the production of C3dg from iC3b in the eye.

[0094] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the production of C3c from iC3b in the eye.

[0095] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing the level of C3b α-chain in the eye.

[0096] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the hydrolysis of peptide substrates in the eye.

[0097] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing proteolysis of macromolecular protein substrates in the eye.

[0098] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing the level or function of the membrane attack complex (MAC) in the eye.

[0099] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing hemolysis observed in the eye.

[0100] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing cleavage of C3b in the absence of cofactors in the eye, e.g., in a cofactor-independent manner.

[0101] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing cleavage of C4b in the absence of cofactors in the eye, e.g., in a cofactor-independent manner.

[0102] The CFI fusion constructs disclosed herein can be used for ocular therapy of a subject. As used herein, a subject includes any mammalian subject, and includes primates, rodents, livestock, zoo animals, and pets. In some embodiments, the mammalian subject is a human subject. In some embodiments, the mammalian subject is a non-human primate.

[0103] B. Treatment of eye diseases

[0104] The CFI fusion constructs provided herein can be used to treat an ocular condition in a subject. In some embodiments, provided herein is a method of treating an ocular condition in a subject in need thereof, the method comprising administering to the subject an intraocularly a therapeutically effective amount of any CFI fusion construct provided herein or a pharmaceutical composition thereof at a dose of about 0.1 mg per eye to about 10 mg per eye and / or at a frequency of about every 1 month to about 4 months.

[0105] In some embodiments, the ocular disorder is characterized by a deficiency of CFI. In some embodiments, the ocular disorder is characterized by a dysregulation of the complement system.

[0106] In some embodiments, the ocular disorder is characterized by the presence of a dysfunctional CFI gene. In some embodiments, the ocular disorder is characterized by dysregulation of the complement system and low CFI levels.

[0107] In some embodiments, the ocular disorder is selected from the group consisting of diabetic macular edema (DME), diabetic retinopathy, glaucoma, keratoconjunctivitis, neuromyelitis optica spectrum disorder (NMOSD), open-angle glaucoma, polypoidal choroidal vasculopathy, Stargardt's disease, uveitis and vitreoretinopathy, and age-related macular degeneration (AMD).

[0108] In some embodiments, the eye condition is dry AMD. In some embodiments, the eye condition is early dry AMD. In some embodiments, the eye condition is mid-term dry AMD. In some embodiments, the eye condition is late dry AMD. In some embodiments, the eye condition is late dry AMD that has progressed to wet AMD. In some embodiments, the eye condition is late dry AMD that has progressed to geographic atrophy (GA).

[0109] In some embodiments, the eye condition is wet AMD. In some embodiments, the eye condition is early wet AMD. In some embodiments, the eye condition is mid-term wet AMD. In some embodiments, the eye condition is late wet AMD. In some embodiments, the eye condition is late wet AMD that has progressed to neovascular AMD.

[0110] In some embodiments, the eye condition is geographic atrophy (GA). In some embodiments, the eye condition is GA that progresses from dry AMD. In some embodiments, GA is associated with late AMD. In some embodiments, GA is not associated with late AMD.

[0111] In some embodiments, the ocular disorder is caused by one or more mutations in the CFI gene of the subject selected from the group consisting of G119R, L131R, V152M, G162D, R187Y, R187T, T203I, A240G, A258T, G287R, A300T, R317W, R339Q, V412M, P553S, K441R, R339Ter, R317Q, G261D, R187Q, and R187Ter.

[0112] In some embodiments, the ocular disorder is caused by one or more mutations in the CFI gene of the subject, the mutations selected from the group consisting of Phe13Val, Val20Ile, His40Arg, Cys43Phe, Asp44Asn, Pro50Ala, Cys54Arg, Cys54*, Ile55Phe, Pro64Leu, Asn70Thr, Thr72Ser, Phe82Ser, Pro83Gln, Ser90Asn, Cys106Arg, Thr107Ala, Glu109Ala, His118Arg, Gly119Arg, Gly125Arg, Val12 A rg187*, Arg187Gln, Gly188Ala, Phe198Leu, Arg202Ile, Thr203Ile, Tyr206Asn, Gln217His, Ser221Tyr, Asp224Asn, Cys229Arg, Val230Met, Val230G A la300Thr, Glu303Lys, Glu305*, Ile306Val, Ile306Ser, Asp310Glu, Arg317Trp, Arg317Gln, Cys327Arg, Gly328Arg, Arg339*, Arg339Gln, Ile340Thr , Gly342Glu, Arg345Gln, Gly349Arg, Val355Met, Ala356Pro, Ile357Met, Gly362Ala, Tyr369Ser, Trp374Cys, Arg389His, Trp399Arg, Pro402Ser, Asp 403Asn, Arg406Cys, Arg406His, Tyr411*, Val412Met, Ile416Leu, His418Leu, Gly424Asp, Asp429Glu, Ala431Thr, Ile433Val, Ile433Thr, Lys441Arg,Arg448Cys, Ala452Ser, Trp456Leu, Tyr459Ser, Gln462His, Cys467Arg, Arg474*, Arg474Gln, Asp477His, Gly487Cys, Ile492Leu, Gly500Arg, Arg502Cys, Gly512Ser, Asp519Asn, Lys522Thr, Asp524Val, Met532Val, Asn536Lys, Trp541*, Gly542Ser, Val543Ala, Trp546*, Glu548Gln, Pro553Ser, Glu554Val, Ser570Thr, Ile578Thr, and Gln580*.

[0113] In some embodiments, the ocular disorder is caused by one or more of a group of mutations in a subject with GA, the group of mutations consisting of: C2, CFB, CFH, and C3 mutations.

[0114] C. Combination therapy

[0115] Administration of any of the therapeutic CFI fusion constructs provided herein can be monotherapy, or can be combined with any other known drug or treatment. Other known drugs or treatments can be used for conditions associated with dysregulation of the complement system, or can be associated with CFI deficiency.

[0116] D. Application

[0117] The CFI fusion constructs described herein are delivered to the eye as polypeptide-based therapeutics.

[0118] Such treatment as contemplated herein comprises administering a CFI fusion construct of the present disclosure to the eye. Accordingly, provided herein are pharmaceutical compositions comprising a CFI fusion construct of the present disclosure.

[0119] Ocular administration of the therapeutic CFI fusion constructs described herein can be intraorbital, periocular, intraocular, intravitreal, or subretinal. Administration of the fusion constructs can be performed with any suitable excipient, carrier, or other agent to provide suitable or improved tolerability, transfer, delivery, etc. In an exemplary embodiment, administration of the CFI fusion constructs described herein is intravitreal administration.

[0120] In some embodiments, the CFI fusion construct can be expressed as an inactive single-chain precursor protein and activated by furin (a serine protease). Furin is an endoprotease that cuts CFI at the conserved RRKR sequence (also known as the furin recognition sequence) to produce a heavy chain and a light chain connected by a disulfide bond. The mature double-chain protein processed by furin is the activated form of the CFI protein. As generally contemplated herein, the CFI fusion construct described herein is delivered in an activated double-chain form. However, in some cases, the inactive CFI fusion construct can be delivered in an inactive single-chain form. In some embodiments, the delivered form includes both a single-chain inactive form and a double-chain active form.

[0121] E. Drug administration

[0122] In embodiments provided herein, a CFI fusion construct described herein is administered to one or both eyes of a subject in need thereof at a dose of about 0.1 mg per eye to about 10 mg per eye and / or at a frequency of about every 4 to 16 weeks.

[0123] The optimal dosage ranges and frequencies of the present disclosure are expected to maintain baseline CFI levels above the levels in subjects with an ocular disease (e.g., AMD) within 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150 days following ocular injection of a CFI fusion construct.

[0124] In some embodiments, after intraocular administration of the fusion CFI construct, the baseline CFI level of a subject with an ocular disease remains higher than the baseline CFI level of a reference subject with the same ocular disease who did not receive the injection. In some embodiments, such CFI levels are maintained for at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150 days after CFI-HSA injection.

[0125] In some embodiments, a CFI fusion construct described herein is administered to an eye of a subject in need thereof at a dose of about 0.1 mg per eye to about 10 mg per eye and at a frequency of about every 4 to 16 weeks.

[0126] In some embodiments, the CFI fusion construct is administered to one eye. In some embodiments, the CFI fusion construct is administered to both eyes. In some embodiments, the CFI fusion construct is administered to both eyes simultaneously. In some embodiments, the CFI fusion construct is administered to both eyes sequentially.

[0127] Exemplary dosages of the CFI fusion constructs described herein are from about 0.1 mg per eye to about 10 mg per eye. In some embodiments, the dosages of the CFI fusion constructs described herein are from about 0.5 mg per eye to about 10 mg per eye, from about 1 mg per eye to about 10 mg per eye, from about 1.5 mg per eye to about 10 mg per eye, from about 2 mg per eye to about 10 mg per eye, from about 2.5 mg per eye to about 10 mg per eye, from about 3 mg per eye to about 10 mg per eye, from about 3.5 mg per eye to about 10 mg per eye, from about 4 mg per eye to about 10 mg per eye, from about 4.5 mg per eye to about 10 mg per eye, g per eye, about 5 mg per eye to about 10 mg per eye, about 5.5 mg per eye to about 10 mg per eye, about 6 mg per eye to about 10 mg per eye, about 6.5 mg per eye to about 10 mg per eye, about 7 mg per eye to about 10 mg per eye, about 7.5 mg per eye to about 10 mg per eye, about 8 mg per eye to about 10 mg per eye, about 8.5 mg per eye to about 10 mg per eye, about 9 mg per eye to about 10 mg per eye, or about 9.5 mg per eye to about 10 mg per eye.

[0128] In some embodiments, administration of a CFI fusion construct described herein is intraocular (e.g., intravitreal) administration at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye.

[0129] In some embodiments, the frequency of administration of the CFI fusion construct is about every 4 weeks to about every 16 weeks. In some embodiments, the administration of the CFI fusion construct is intravitreally administered and is about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0130] In some embodiments, administration of a therapeutic CFI fusion construct described herein is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 4 weeks.

[0131] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 5 weeks.

[0132] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 6 weeks.

[0133] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 7 weeks.

[0134] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 8 weeks.

[0135] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 9 weeks.

[0136] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 10 weeks.

[0137] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 11 weeks.

[0138] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 12 weeks.

[0139] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 13 weeks.

[0140] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 14 weeks.

[0141] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 15 weeks.

[0142] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye, about 0.5 mg per eye, about 1 mg per eye, about 1.5 mg per eye, about 2 mg per eye, about 2.5 mg per eye, about 3 mg per eye, about 3.5 mg per eye, about 4 mg per eye, about 4.5 mg per eye, about 5 mg per eye, about 5.5 mg per eye, about 6 mg per eye, about 6.5 mg per eye, about 7 mg per eye, about 7.5 mg per eye, about 8 mg per eye, about 8.5 mg per eye, about 9 mg per eye, about 9.5 mg per eye, or about 10 mg per eye, at a frequency of about every 16 weeks.

[0143] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0144] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0145] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 1.0 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0146] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 1.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0147] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 2 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0148] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 2.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0149] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 3 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0150] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 3.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0151] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 4 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0152] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 4.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0153] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0154] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 5.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0155] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 6 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0156] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 6.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0157] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 7 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0158] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 7.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0159] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 8 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0160] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 8.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0161] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 9 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0162] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 9.5 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0163] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 10 mg per eye at a frequency of about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, about every 13 weeks, about every 14 weeks, about every 15 weeks, or about every 16 weeks.

[0164] F. Preparation

[0165] Pharmaceutical compositions containing the CFI fusion constructs of the present disclosure can be formulated in any conventional manner suitable for ocular administration by mixing a selected amount of the polypeptide with one or more physiologically acceptable carriers or excipients for use in the treatments provided herein. The choice of carrier or excipient is within the skill of the administration profession and may depend on a variety of parameters. These parameters include, for example, the mode of administration and the condition being treated. The pharmaceutical compositions provided herein may be formulated for single dose (direct) administration or for dilution or other modification. The concentration of the compound in the formulation is effective to deliver an amount effective for the intended treatment when administered. Typically, the composition is formulated for single dose administration, but this is not required.

[0166] In some exemplary embodiments, the fusion construct can be concentrated to at least 150 mg / mL. In some embodiments, the fusion construct can be dosed to at least 7.5 mg in 50 uL. In some embodiments, the fusion construct can be dosed to about 25 ul to about 150 ul, about 30 ul to about 150 ul, about 35 ul to about 150 ul, about 40 ul to about 150 ul, about 45 ul to about 150 ul, about 50 ul to about 150 ul, about 55 ul to about 150 ul, about 60 ul to about 150 ul, about 65 ul to about 150 ul, about 70 ul to about 150 ul, about 75 ul to about 150 ul, about 80 ul to about 150 ul, about 85 ul to about 150 ul , at least 7.5 mg in about 90 ul to about 150 ul, about 95 ul to about 150 ul, about 100 ul to about 150 ul, about 105 ul to about 150 ul, about 110 ul to about 150 ul, about 115 ul to about 150 ul, about 120 ul to about 150 ul, about 125 ul to about 150 ul, about 130 ul to about 150 ul, about 135 ul to about 150 ul, about 140 ul to about 150 ul, about 145 ul to about 150 ul, or about 25 ul to about 150 ul.

[0167] In some exemplary embodiments, the fusion construct can be dosed in 25 uL at least 7.5 mg, 30 uL at least 7.5 mg, 35 uL at least 7.5 mg, 40 uL at least 7.5 mg, 45 uL at least 7.5 mg, 50 uL at least 7.5 mg, 55 uL at least 7.5 mg, 60 uL at least 7.5 mg, 65 uL at least 7.5 mg, 70 uL at least 7.5 mg, 75 uL at least 7.5 mg, 80 uL at least 7.5 mg, 85 uL at least 7.5 mg, 7.5 mg, at least 7.5 mg in 90uL, at least 7.5 mg in 95uL, at least 7.5 mg in 100uL, at least 7.5 mg in 105uL, at least 7.5 mg in 110uL, at least 7.5 mg in 115uL, at least 7.5 mg in 120uL, at least 7.5 mg in 125uL, at least 7.5 mg in 130uL, at least 7.5 mg in 135uL, at least 7.5 mg in 140uL, at least 7.5 mg in 145uL or at least 7.5 mg in 150uL.

[0168] In some exemplary embodiments, the fusion construct is dosed at about 5 to 10 mg in 25uL, 30uL, 35uL, 40uL, 45uL, 50uL, 55uL, 60uL, 65uL, 70uL, 75uL, 80uL, 85uL, 90uL, 95uL, 100uL, 105uL, 110uL, 115uL, 120uL, 125uL, 130uL, 135uL, 140uL, 145uL, or 150uL.

[0169] G. Pharmaceutical Compositions

[0170] The present disclosure also provides a pharmaceutical composition comprising any of the CFI fusion constructs disclosed herein and optionally a pharmaceutically acceptable excipient or carrier suitable for ocular administration. In some embodiments, the pharmaceutical composition is sterile. The pharmaceutical composition can be formulated to be compatible with its intended route of administration. In some embodiments, the pharmaceutical composition of the present disclosure is suitable for administration to human subjects or other non-human primates. In an exemplary embodiment, the pharmaceutical composition is formulated for intravitreal administration.

[0171] H. Kits and Articles of Manufacture of Therapeutic CFI Fusion Constructs

[0172] The present disclosure also provides a kit or article comprising any of the CFI fusion constructs disclosed herein or any pharmaceutical composition disclosed herein. In some embodiments, the kit may also include instructional materials for performing any of the methods of administration and intraocular administration disclosed herein. In some embodiments, the kit may also include a sterile container or vial for containing the fusion constructs and / or pharmaceutical compositions disclosed herein. In some embodiments, the kit may also include a sterile delivery device for administering the fusion constructs and / or pharmaceutical compositions disclosed herein. In some embodiments, the article comprises any pharmaceutical composition disclosed herein.

[0173] Example

[0174] Example 1: CFI-HSA expression, purification, activation and in vitro sialylation

[0175] Overview

[0176] With respect to Example 1, reference to CFI-HSA refers to human serum albumin (SEQ ID NO: 21) fused to the N-terminus of wild-type CFI.

[0177] Wild-type CFI-HSA protein was expressed in Chinese hamster ovary (CHO) cells, purified using anti-albumin affinity purification, activated with furin, and purified by fractionation column. The activated CFI-HSA protein was sialylated in vitro to increase the total sialylation of CFI-HSA. Finally, the sialylated protein was purified using anti-albumin affinity purification and refined by size exclusion column chromatography.

[0178] Express

[0179] The CFI-HSA gene (SEQ ID NO:21) was synthesized (ThermoFisher Scientific, Geneart, Regensburg, Germany) with human serum albumin at the amino terminus of the CFI protein. The protein was prepared with a signal sequence of SEQ ID NO:2, which was removed during expression. The amino terminal albumin tag was connected to the CFI gene via a linker (SEQ ID NO:6). The gene for CFI-HSA was inserted into an expression vector (LakePharma, Hayward, CA) using standard molecular biology techniques. The resulting plasmid DNA was transformed into Escherichia coli (E. coli). The transfected E. coli was grown in 200 mL LB medium to express the plasmid DNA and harvested using standard techniques. The plasmid DNA was run on an agarose gel for quality assessment and the sequence was confirmed before transfection.

[0180] 1.0 L of suspended TunaCHO TM Cells were seeded in shake flasks and expanded using serum-free chemically defined medium. On the day of transfection, expanded cells were inoculated into new flasks with fresh medium. Plasmid DNA was transiently transfected into CHO cells using Lipofectamine 2000 (ThermoFisher Scientific). Cells were maintained as fed-batch cultures until the end of the production run. The protein was grown at 37°C at 125 RMP and 8% CO 2 The cells were centrifuged and the supernatant was collected to purify the secreted CFI-HSA at the end of the 14-day expression.

[0181] purification

[0182] The supernatant containing the expressed CFI-HSA protein was passed through CaptureSelect TMA 10 mL gravity flow column of human albumin affinity matrix (ThermoFisher Scientific) was used. The column-bound protein was washed with 10 column volumes of 20 mM sodium phosphate buffer. The bound CFI-HSA protein was eluted in two steps: first, with 3 column volumes of 20 mM Tris-HCl (pH 7.0) buffer and 2 M MgCl 2 Elution, second step, eluted with 3 column volumes of 20 mM citric acid (pH 3.0). Collect the eluates from both steps 1 and 2 in 5 mL fractions. Neutralize each fraction of the step 2 eluate with 10% neutralization buffer (1.5 M tris-HCL, pH 7.4). Analyze all fractions by reducing and non-reducing SDS-PAGE electrophoresis and analyze by SimplyBlue TM SafeStain (ThermoFisher Scientific) was used to visualize the bands. CFI-HSA ran as a 130 kDa band on the non-reducing gel and as 102 kDa and 28 kDa bands on the reducing gel. Fractions with the greatest CFI-HSA concentration and purity were pooled for further processing.

[0183] Furin activation

[0184] The purified CFI-HSA was prepared by incubating 4 μg of recombinant furin / mg of purified CFI-HSA at 30°C in Tris-NaCl (tris-buffered saline), 2.5 mM CaCl 2 CFI-HSA was cleaved to produce the protein in an activated form by incubation in 0.5% CHAPS for 18 hours. The CFI-HSA protein concentration was maintained at 1.4 mg / mL. This resulted in more than 90% of the protein being activated. The activated protein was separated from the inactivated CFI-HSA and other proteins by size exclusion chromatography. The HPLC-MS / MS was performed using a HiLoad 16 / 600 Superdex 200 column (GE Healthcare Life Sciences) and phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 2 mM KH 2 PO 4 Size exclusion chromatography (SEC) was performed with 5% paraformaldehyde (pH 7.4) as the mobile phase. The collected fractions were analyzed by CE-SDS (LabChip GXII, Perkin Elmer). The fractions containing the target protein were combined and analyzed by SE-UPLC.

[0185] In vitro sialylation

[0186] The activated CFI-HSA protein was sialylated in vitro. Briefly, sialylation was performed in a two-step enzymatic reaction. First, galactosyltransferase (GalT1) and CFI-HSA were used at a molar ratio of 1:200 in the presence of 10 mM UDP-galactose, 5 mM MnCl, and 1% daptomycin. 2 The galactosylation reaction of CFI-HSA was carried out in 200 μL volume in 100 mM MES (pH 6.5) buffer. TM Galactosylated CFI-HSA was purified from the reaction mixture by human albumin affinity chromatography. Next, α2,6-sialyltransferase and purified CFI-HSA were used at a molar ratio of 1:50 in the presence of 80 μM alkaline phosphatase, 6.1 mM CMP-NANA, 10 mM ZnCl 2 The sialylation reaction was carried out in 250 μL volume in 200 mM MES buffer (pH 6.5) at 37°C for 1 hour. TM Human albumin affinity chromatography was used to purify sialylated CFI-HSA protein from the reaction mixture. The level and characteristics of sialic acid chains on CFI-HSA were determined by using the Agilent / Prozyme Analytical Services "GS-SAP Method for Total Sialic Acid Quantification (Agilent GS48)" and mass spectrophotometric (MS) analysis (Lake Pharma Analytical Services), as described in further detail below.

[0187] In brief, total sialic acid is quantitatively carried out by mixing 20 μ L of each sample with 10 μ L of release reagent in 96 well plates. The reaction mixture is incubated at 80 ℃ for 2 hours. The sample is cooled to room temperature and 10 μ L of labeling reagent is added to each sample to further incubate for 3 hours at 50 ℃. The sample is cooled to room temperature again and 160 μ L of deionized (dI) water is added to make the total volume reach 200 μ L. 10 μ L of sample is injected into Agilent UHPLC Poroshell C18 column, at 30 ℃ in 4% methanol, 8% acetonitrile aqueous solution (line A1) and 100% ACN (line B1) with a flow rate of 0.4mL / minute. The peak value at 373 / 448nm wavelength is recorded. The standard curve of total peak area contrast picomoles (pmol) sialic acid is produced by running the 1pmol-2000pmol NANA (N-acetylneuraminic acid, Neu5Ac) provided by the test kit on the same column. The total sialic acid of each sample was quantified by comparing the peak area of ​​the sample with the standard curve. The sialylation obtained is summarized in Table 1.1 below.

[0188] Table 1.1: Sialylation assay results

[0189] protein Sialic acid (Neu5Ac) pmol / ug protein Recombinant CFI-HSA 35±0.7 In vitro sialylation of recombinant CFI-HSA 69±2.2 Bovine fetuin control 222±2.6

[0190] Mass spectrometry analysis was performed by a standard trypsin Q-TOF mass spectrometer. In brief, all samples were treated, reduced and alkylated with DTT and iodoacetamide, followed by trypsin digestion. The digested samples were analyzed by a Waters ACQUITY UPLC coupled to a Xevo G2-XS-QTOF mass spectrometer using a protein BEH C18 column. The analyses performed are summarized in Table 1.2 below.

[0191] Table 1.2: Peptide analysis results

[0192]

[0193] refined

[0194] The purified CFI-HSA protein was subjected to size exclusion chromatography (SEC) using a HiLoad 16 / 600 Superdex 200 column (GE Healthcare Life Sciences) and phosphate buffered saline as the mobile phase. The collected fractions were analyzed by CE-SDS (LabChip GXII, Perkin Elmer). The fractions containing the target protein were combined to a concentration of 5 mg / mL, and the samples were snap frozen and stored at -80°C.

[0195] N-terminal albumin fusion provides solubility and promotes activation of CFI-HSA .

[0196] Comparison of activation was performed between CFI-HSA and wild-type CFI (WT-CFI) without albumin or other fusion tags. The gene construct of WT-CFI was expressed essentially as described above for CFI-HSA. Recombinant WT-CFI protein showed moderate purity by reducing SDS-PAGE, however, a large amount of high molecular weight species (HMWS) and aggregates were observed under reducing and non-reducing conditions. For CFI-HSA with an N-terminal HSA tag added, TunaCHO TMCells were transiently expressed and subsequently purified as described above, showing no HMWS or aggregates on reduced and non-reduced SDS-PAGE. Activation of purified recombinant CFI with furin resulted in further increases in aggregates and HMWS with almost complete polydispersity. In addition, activated CFI was substantially not observed by reduced SDS-PAGE. In contrast, addition of furin activated CFI-HSA almost completely and effectively under the same conditions, and the CFI-HSA protein remained as a monomer under non-reducing conditions, with no evidence of aggregates and HMWS. When compared to CFI lacking any fusion tag, the N-terminal HSA tag has significant and unexpected benefits for maintaining solubility, monodispersity, and effective furin activation.

[0197] Example 2: Half-life of CFI-HSA in the vitreous humor of non-human primates

[0198] For Example 2, reference to CFI-HSA refers to human serum albumin (SEQ ID NO: 21) fused to the N-terminus of wild-type CFI.

[0199] Pharmacokinetics and pharmacodynamics of CFI-HSA after intravitreal injection

[0200] After intravitreal administration to six AGMs, ocular pharmacokinetics of the N-terminal albumin fusion of wild-type CFI-HSA were examined. Six animals were divided into two groups and treated at two dose levels: one group received a single intravitreal injection of 500 μg CFI-HSA (right eye, OD, N=3), and the other group received a single intravitreal injection of 250 μg CFI-HSA (right eye, OD, N=3). The left eye (OS) of all six animals was injected with an equal volume of 100 μL sterile injection PBS as a vehicle control. Non-terminal vitreous humor samples (100 μL) were collected on days 1, 7, 14, 21, and 28 after administration. The vitreous humor CFI-HSA drug concentration was determined using a quantitative electrochemiluminescence (ECL) antigen assay, which is optimized for measuring CFI-HSA in the vitreous humor of AGMs. The assay uses anti-CFI antibody (clone OX21, LS Bio, Seattle WA) coated at 2 μg / mL on Meso Scale Discovery (MSD, Rockville, MA) assay plates to capture CFI-HSA levels. The captured CFI-HSA is detected at 0.5 μg / mL with a sheep polyclonal anti-HSA antibody (Abcam, Cambridge, MA) conjugated to SULFO-TAG, which emits light [electrochemiluminescence (ECL)] when an electric potential is applied. ECL relative light units (RLU) were measured on a SECTOR S 600 reader and unknown CFI-HSA concentrations in the vitreous humor were interpolated from a standard curve ranging from 0.05 μg / mL to 40 μg / mL Factor I-HSA. The data are provided in Table 2.1.

[0201] Table 2.1 CFI-HSA levels on days 1, 7, 14, 21 and 28 after intravitreal administration

[0202]

[0203] *BLQ: measurement result below the limit of quantitation of the assay

[0204] Non-compartmental analysis yielded apparent ocular terminal half-lives of 3.6 and 4.1 days for the 250 μg and 500 μg dose levels, respectively.

[0205] Table 2.2. Estimated PK parameters of CFI-HSA in the vitreous humor of African green monkeys

[0206]

[0207]

[0208] Complement component 3a (C3a) levels in the vitreous humor were determined by ELISA using the Quidel kit for C3a ELISA for the 250 μg and 500 μg doses (Figure 2). The CFI-HSA fusion construct reduced ocular C3a levels in a dose-dependent manner for up to 7 days after intraocular injection. The increase in C3b degradation by CFI-HSA reduced complex formation between C3b and Bb, which resulted in a reduction in the cleavage of C3 into C3a and C3b via the amplification loop of the alternative pathway.

[0209] After intravitreal administration to six AGMs, ocular pharmacokinetics of N-terminal albumin fusion of wild-type CFI-HSA were examined. Six animals were divided into two groups and treated at two dose levels: one group received a single intravitreal injection of 0.25 mg CFI-HSA (OU, N=4), 1.0 mg CFI-HSA (OU, N=4), and the control group received a single intravitreal injection of vehicle (OU, N=4). All 12 animals were injected with an equal volume of 100 μL test article or vehicle control in the eyes (OU). Animals received ocular examinations on days 0, 1, 7 or 8, 14 or 15, 21 and 28 after administration (see Example 4). Animals were euthanized on days 1, 7, 14 and 28 for PK-PD analysis and histopathology. Quantitative electrochemiluminescence (ECL) antigen assay was used to determine the vitreous CFI-HSA drug concentration, which was optimized for measuring CFI-HSA in the vitreous humor of AGMs as described above. The estimated terminal vitreous half-life measurements for the 0.25 mg and 1.0 mg doses were 4 days and 4.4 days, respectively ( Figure 3A ).

[0210] For the 1.0 mg dose, the levels of complement component 3b (C3b) and cleaved fragment iC3b in the vitreous humor were determined by chemiluminescent Western blotting using anti-C3 antibody and by densitometric quantification ( Figure 2B ). The CFI-HSA fusion construct reduced ocular iC3b levels for up to 14 days after intraocular injection. The increase in C3b degradation by CFI-HSA is expressed as the C3b / iC3b ratio on days 1, 7, and 14, which showed a significant increase in cleavage in eyes injected with 1.0 mg CFI-HSA compared to vehicle controls ( Figure 2B ).

[0211] Example 3: Pharmacokinetic Modeling of Intravitreal Injection of CFI-HSA for Human Administration

[0212] The half-life of CFI-HSA following intravitreal injection of CFI-HSA was measured over time in nonhuman primates. A half-life of approximately 4 days was found to maintain dose proportionality across 3 dose levels in AGM, as Figure 3A In addition, the cleavage activity of CFI-HSA for up to 14 days was detected by Western blotting ( Figure 2B ).

[0213] Based on the AGM data, a vitreous PK model for human dosing was generated, such as Figure 3BWithout being bound by theory or mechanism, it is expected that intravitreal injection of 1 mg per eye of CFI-HSA will maintain a baseline CFI level above the level of AMD patients within 60 days after injection. It is expected that intravitreal injection of 10 mg per eye of CFI-HSA will maintain a baseline CFI level above the level of AMD patients within 90 days after injection.

[0214] Example 4: Safety profile of intravitreal injection of CFI-HSA in non-human primates .

[0215] The safety profile of intravitreal injections of CFI-HSA was determined in nonhuman primates. After injection, clinical scores of AGM were assessed by slit-lamp testing on days 0, 1, 7 or 8, 14 or 15, 21, and 28 or until termination. Clinical scores were generated over time using slit-lamp testing after intravitreal injection of CFI-HSA, as Figure 4A The intraocular pressure of the AGM subjects after CFI-HSA injection was also evaluated. The intraocular pressure of the injected AGM subjects was measured over time, as shown in FIG. Figure 4B The intraocular pressure of each injected AGM subject was found to be within the baseline variation and thus stable. Finally, the retinal thickness of the AGM subjects was assessed after CFI-HSA injection. The retinal thickness of the injected AGM subjects was measured over time, as shown in FIG. Figure 4C The retinal thickness of each AGM injected subject was found to be within the baseline variation and thus stable.

Claims

1. A method of treating or preventing an ocular condition in a human subject in need thereof, comprising: administering to the subject a fusion construct comprising (a) complement factor I (CFI) comprising the amino acid sequence of SEQ ID NO.5 or a variant thereof, and (b) a binding partner, wherein the route of administration is intraocular, and wherein the fusion construct is administered in the following manner: (a) a dose of about 0.1 mg per eye to about 10 mg per eye; and / or (b) A frequency of about every 1 month to about 4 months.

2. The method of claim 1, wherein the variant of CFI has at least 70%, at least 80%, at least 90% or at least 95% sequence identity with the amino acid sequence of SEQ ID NO.

5.

3. The method of claim 2, wherein the variant of CFI comprises CFI of SEQ ID NO: 5 comprising one or more of substitutions T377G, N422K, E457G, and N531G.

4. The method of claim 2, wherein the variant of CFI comprises CFI of SEQ ID NO: 5 comprising substitutions N422K, E457G, and N531G.

5. The method of claim 2, wherein the variant of CFI comprises CFI of SEQ ID NO: 5 comprising substitutions T377G, E457G, and N531G.

6. The method of claim 1, wherein the fusion protein comprises complement factor I (CFI), and the complement factor I comprises the amino acid sequence of SEQ ID NO.

5.

7. The method according to any one of claims 1 to 6, wherein the binding partner is serum albumin or a variant thereof comprising the amino acid sequence of SEQ ID NO.

7.

8. The method of claim 7, wherein the variant of serum albumin has at least 70%, at least 80%, at least 90% or at least 95% sequence identity with the amino acid sequence of SEQ ID NO.

7.

9. The method according to any one of claims 1 to 8, wherein the fusion construct has at least 70%, at least 80%, at least 90% or at least 95% sequence identity with the amino acid sequence of SEQ ID NO.

21.

10. The method according to any one of claims 1 to 9, wherein the fusion construct comprises the amino acid sequence of SEQ ID NO.

21.

11. The method according to any one of claims 1 to 6, wherein the binding partner is a cofactor of CFI.

12. The method of claim 11, wherein the binding partner comprises a factor H cofactor comprising the amino acid sequence of SEQ ID NO: 22 or SEQ ID NO: 23, a domain thereof, or a portion of a domain thereof.

13. The method of claim 11, wherein the binding partner comprises CR1, a domain thereof, or a portion of a domain thereof.

14. The method according to any one of claims 11 to 13, comprising serum albumin comprising the amino acid sequence of SEQ ID NO. 7 or a variant thereof.

15. The method of claim 14, wherein the variant of serum albumin has at least 70%, at least 80%, at least 90% or at least 95% sequence identity with the amino acid sequence of SEQ ID NO.

7.

16. The method of any one of claims 1 to 15, wherein the ocular condition is dry age-related macular degeneration (AMD).

17. The method of any one of claims 1 to 15, wherein the ocular condition is wet AMD.

18. The method of any one of claims 1 to 15, wherein the ocular disorder is geographic atrophy (GA).

19. The method of any one of claims 1 to 18, wherein the subject exhibits geographic atrophy (GA).

20. The method of any one of claims 1 to 19, wherein the human subject carries one or more mutations of CFI selected from the group consisting of G119R, L131R, V152M, G162D, R187Y, R187T, T203I, A240G, A258T, G287R, A300T, R317W, R339Q, V412M, P553S, K441R, R339Ter, R317Q, G261D, R187Q, and R187Ter.

21. The method of any one of claims 1 to 20, wherein the dosage is about 0.1 mg per eye to about 10 mg per eye, about 0.5 mg per eye to about 10 mg per eye, about 1 mg per eye to about 10 mg per eye, about 1.5 mg per eye to about 10 mg per eye, about 2 mg per eye to about 10 mg per eye, about 2.5 mg per eye to about 10 mg per eye, about 3 mg per eye to about 10 mg per eye, about 3.5 mg per eye to about 10 mg per eye, about 4 mg per eye to about 10 mg per eye, about 4. 5 mg per eye to about 10 mg per eye, about 5 mg per eye to about 10 mg per eye, about 5.5 mg per eye to about 10 mg per eye, about 6 mg per eye to about 10 mg per eye, about 6.5 mg per eye to about 10 mg per eye, about 7 mg per eye to about 10 mg per eye, about 7.5 mg per eye to about 10 mg per eye, about 8 mg per eye to about 10 mg per eye, about 8.5 mg per eye to about 10 mg per eye, about 9 mg per eye to about 10 mg per eye, or about 9.5 mg per eye to about 10 mg per eye.

22. The method of any one of claims 1 to 21, wherein the dose is about 0.1 mg per eye, 0.5 mg per eye, 1 mg per eye, 5 mg per eye, or 10 mg per eye.

23. The method of any one of claims 1 to 22, wherein the frequency is about every 1 month.

24. The method of any one of claims 1 to 22, wherein the frequency is about every 2 months, 3 months, or 4 months.

25. The method of any one of claims 1 to 24, wherein the fusion construct is administered to one eye.

26. The method of any one of claims 1 to 24, wherein the fusion construct is administered to both eyes.

27. The method of any one of claims 1 to 24, wherein the fusion construct is administered sequentially to both eyes.

28. The method of any one of claims 1 to 24, wherein the fusion construct is administered to both eyes simultaneously.

29. The method of any one of claims 1 to 28, wherein the subject's intraocular pressure is stable after treatment.

30. The method of any one of claims 1 to 29, wherein the subject's retinal thickness is stable following treatment.

31. The method of any one of claims 1 to 30, wherein following administration, the fusion construct exhibits an activity in the subject that is at least equivalent to the activity exhibited by endogenous CFI.

32. The method of any one of claims 1 to 30, wherein after administration, the baseline CFI level remains above the baseline CFI level of a reference subject who did not receive the injection.

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