Complement factor I dosing regimen for treating ocular diseases

Abstract

Provided herein is a dosing regimen for the treatment of ocular conditions, such as age-related macular degeneration (AMD), comprising administering to a subject in need thereof a fusion construct comprising complement factor I (CFI) and a binding partner at a particular dose and frequency.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 63 / 338,392, filed May 4, 2022, the entire contents of which are incorporated herein by reference.

[0002] (Reference to Electronic Sequence Listing) 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 technology]

[0003] The complement system includes the classical, lectin, and alternative pathways and is tightly controlled by several regulatory factors. Complement Factor I (CFI) is one such regulatory factor, which acts to regulate the complement system by cleaving and thereby inactivating C4b and C3b proteins. Such cleavage results in the inhibition of the classical, lectin, and alternative pathways, respectively, thus ultimately preventing the assembly of C3 and C5 convertase enzymes. CFI is encoded as a proenzyme, which is then activated by proteolytic cleavage into a heterodimeric glycoprotein with a heavy chain (A chain) and a light chain (B chain) connected by a disulfide bond. CFI is post-translationally processed into its active form by furin to generate a two-chain mature protein.

[0004] Dysregulation of CFI, mutations and dysfunctions of CFI (i.e., genetic variants), or CFI deficiency are associated with diseases involving the complement system. Dysregulation or overactivation of the complement system has been associated with eye diseases such as Age-related Macular Degeneration (AMD). AMD progression leads to damage of the retina macula, atrophy of the retinal pigment epithelium (RPE), and loss of photoreceptors, causing blurred vision and eventual vision loss. Carrying rare genetic variants of CFI and / or low CFI levels has been associated with AMD and disease progression. According to the National Eye Institute of the National Institutes of Health (NIH), AMD is the leading cause of vision loss in adults, affecting more than 190,000,000 people 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 in the back of the eye, damaging the macula. Dry AMD is characterized by thinning of the macula and may progress to geographic atrophy (GA), which is characterized by progressive and irreversible loss of retinal tissue. GA is the leading cause of blindness, affecting more than 5,000,000 people worldwide. There is a need for a method to regulate or inhibit the complement system to maintain regulatory balance for the treatment of eye 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

[0005] In one aspect, provided herein is 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 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 the fusion construct is administered at a dose of about 0.1 to about 10 mg / eye and / or at a frequency of about every 1 to 4 months. In some embodiments, the binding partner is serum albumin comprising the amino acid sequence of SEQ ID NO:7, or a variant thereof. In some embodiments, the variant of CFI 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, the variant of CFI comprises a CFI of SEQ ID NO:5 comprising one or more of the following substitutions: T377G, N422K, E457G, and N531G, or comprises a CFI of SEQ ID NO:5 comprising the following substitutions: N422K, E457G, and N531G, or comprises a CFI of SEQ ID NO:5 comprising the following 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 a domain thereof. In some embodiments, the binding partner comprises CR1, a domain thereof, or a portion of a domain thereof. The CFI-FH fusion protein can further 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, e.g., 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 in 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, R187Q, and R187Ter. In some embodiments, the ocular condition is Phe13Val, Val20Ile, His40Arg, Cys43Phe, Asp44Asn, Pro50Ala, Cys54Arg, Cys54. * , Ile55Phe, Pro64Leu, Asn70Thr, Thr72Ser, Phe82Ser, Pro83Gln, Ser90Asn, Cys106Arg, Thr107Ala, Glu109 Ala, His118Arg, Gly119Arg, Gly125Arg, Val127Ala, Val129Gly, Leu131Arg, Met138Val, Met138Ile, Trp145 * , Asn151Ser, Val152Met, Gly162Asp, Asp164Val, Asn177Ile, His183Arg, Val184Met, Arg187 * , Arg187Gln, Gly188Ala, Phe198Leu, Arg202Ile, Thr203Ile, Tyr206Asn, Gln217His, Ser221Tyr, Asp224Asn, Cys229Arg, Val230Met, Val230Glu, Ala240Gly, Cy s247Gly, Gly248Glu, Asp249Glu, Ala258Thr, Gly261Ser, Gly261Asp, Gly263Val, Lys267Asn, Gly280Asp, Gly287Arg, Glu290Asp, Ala300Thr, Glu303Lys, Glu305 * , Ile306Val, Ile306Ser, Asp310Glu, Arg317Trp, Arg317Gln, Cys327Arg, Gly328Arg, Arg339 *, Arg339Gln, Ile340Thr, Gly342Glu, Arg345Gln, Gly349Arg, Val355Met, Ala356Pro, Ile357Met, Gly362A la, Tyr369Ser, Trp374Cys, Arg389His, Trp399Arg, Pro402Ser, Asp403Asn, Arg406Cys, Arg406His, Tyr411 * , Val412Met, Ile416Leu, His418Leu, Gly424Asp, Asp429Glu, Ala431Thr, Ile433Val, Ile433T hr, 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 frequency is about once a month. In some embodiments, the frequency is about once every 2, 3, 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 the treatment. In some embodiments, the retinal thickness of the subject is stable after the treatment. In some embodiments, the fusion construct exhibits an activity in the subject that is at least equivalent to the activity exhibited by plasma-derived or naturally occurring CFI protein. [Brief description of the drawings]

[0006] [Figure 1] FIG. 1 shows a model of an exemplary CFI-albumin (e.g., serum albumin, e.g., Human Serum Albumin (HSA)) fusion construct comprising serum albumin fused to CFI, where the CFI comprises wild-type CFI. [Figure 2A] FIG. 1 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. [Figure 2B] FIG. 1 is a plot showing the ratio of C3b / iC3b 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 1.0 mg in an AGM primate model. [Figure 3A]1 is a graph depicting vitreous pharmacokinetics (PK) in AGM. Constructs containing CFI-HSA were injected intravitreally at 0.25 mg / eye or 1 mg / eye. The amount of injected construct was measured over time. [Figure 3B] 1 is a graph depicting vitreous PK modeling in humans based on the PK measured for AGM. [Figure 4A] 1 is a graph depicting slit lamp clinical scores for intravitreal injection of CFI-HSA in non-human primates. [Figure 4B] 1 is a graph showing intraocular pressure measured over time in non-human primates following intravitreal injection of CFI-HSA. [Figure 4C] 1 is a graph showing retinal thickness measured over time in non-human primates following intravitreal injection of CFI-HSA. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] The present disclosure provides dosing regimens of fusion constructs comprising CFI (e.g., comprising HSA, Factor H, or CR1) and binding partners useful for regulating complement signaling and amplification in the ocular system. The present disclosure also provides methods of making and using these fusion constructs in treating ocular diseases or conditions associated with complement dysregulation, such as, for example, dry age-related macular degeneration (AMD), wet AMD, and geographic atrophy (GA).

[0008] I. Complement Factor I Fusion Constructs Complement factor I (CFI) fusion constructs are provided herein, such fusion constructs comprising: (a) a wild-type CFI or a mutant thereof; (b) a binding partner.

[0009] 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.

[0010] In some embodiments, the fusion construct comprises: (a) complement factor I comprising the amino acid sequence of SEQ ID NO:5 or a variant thereof; (b) serum albumin comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof.

[0011] In some embodiments, the fusion construct comprises: (a) complement factor I comprising the amino acid sequence of SEQ ID NO:5 or a variant thereof; (b) a factor H cofactor comprising the amino acid sequence of SEQ ID NO: 22 or SEQ ID NO: 23 or a domain thereof; (c) optionally, a serum albumin comprising the amino acid sequence of SEQ ID NO:7 or a variant thereof.

[0012] In some embodiments, the fusion construct comprises: (a) complement factor I comprising the amino acid sequence of SEQ ID NO:5 or a variant thereof; (b) a CR1 cofactor or a domain thereof; (c) optionally, a serum albumin comprising the amino acid sequence of SEQ ID NO:7 or a variant thereof.

[0013] As used herein, "wild type CFI" refers to any naturally occurring full-length or nearly full-length CFI that is not a disease-causing CFI, which may be of any species, with or without a signal sequence.

[0014] 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 (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 (shown in Table 1 below).

[0015] Wild-type CFI comprises a heavy chain and a light chain, which are also referred to as chain A and chain B, respectively. Figure 1 depicts a schematic diagram of CFI showing the two chains.

[0016] Exemplary base molecules used as components of CFI fusion constructs are provided in Table 1. The base molecules in Table 1 were used to generate the CFI fusion constructs disclosed herein. The base molecules in Table 1 may be further moieties of fusion constructs, as described further below.

[0017] Provided herein are fusion constructs that contain variants to CFI that contain one or more modifications with respect to wild-type CFI, referred to herein as "CFI variants." As used herein, a "modification" relative to wild-type CFI includes a deletion of one or more amino acid residues, a deletion of one or more domains, a substitution of one or more amino acid residues, an insertion (i.e., addition) of one or more amino acid residues, an insertion (i.e., addition) of one or more domains, an inversion of one or more domains, and a substitution of one or more domains.

[0018] Mutants of CFI may include those that add additional activity to CFI in one or more of the classical pathway, the lectin pathway, and the alternative pathway. In some embodiments, the modification to wild-type CFI includes 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 swapping of one or more CFI domains. PCT Publication WO / US2021 / 037278 and PCT Publication WO / US2022 / 082179 provide mutants of CFI and are incorporated herein by reference in their entireties.

[0019] In some embodiments, the CFI comprises any one or more domains selected from the Serine Protease Domain (SPD), the Factor I Membrane Attack Complex (FIMAC) domain, the SRCR domain, the Low Density Lipoprotein receptor 1 (LDLr1) domain, and the Low Density Lipoprotein receptor 2 (LDLr2) domain.

[0020] In some embodiments, a CFI variant contains at least one modification corresponding to wild-type human CFI. In some embodiments, a CFI variant contains at least one modification corresponding to wild-type non-human CFI. In some embodiments, a CFI variant contains at least one modification corresponding to wild-type CFI having the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:5.

[0021] In some embodiments, the human CFI is a chimera comprising one or more domains from human CFI, and the human CFI further comprises a substitution of one or more amino acid residues for amino acid residues in a corresponding region from a non-human species CFI. In some embodiments, the non-human species is mouse. In some embodiments, the CFI variant is chimeric, and the modification comprises a substitution of one or more amino acid residues of CFI with amino acid residues from a corresponding region of a non-CFI serine protease. In some embodiments, the non-CFI serine protease is trypsin.

[0022] In some embodiments, the CFI variants comprise an A chain and a B chain, and the CFI variants comprise one or more modifications at the interface between the A and B chains.

[0023] In some embodiments, the CFI variants comprise modifications at any one or more positions corresponding to positions K14, Y20, D26, F29, R35, E38, M220, K221, S250, L304, P305, K306, L307, and S308 of a CFI having an amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI variants comprise one or more of substitutions selected from K14A, Y20A, Y20F, D26A, F29A, R35A, E38A, M220A, K221Q, S250A, S250L, L304G, P305G, K306G, L307G, and S308G, where the positions correspond to positions of a CFI having an amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI mutant comprises one or more of the substitutions M220A, K221Q, and combinations of L304G, P305G, K306G, L307G, and S308G, positions which correspond to positions in a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0024] In some embodiments, a CFI variant comprises one or more modifications in the C-terminal region of the CFI variant, hi some embodiments, a CFI variant comprises modifications 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 a CFI having the amino acid sequence set forth in SEQ ID NO:5. In some embodiments, the CFI mutant comprises one or more substitutions selected from 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, where the positions correspond to those of a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0025] In some embodiments, the CFI variants comprise one or more modifications at one or more N-linked glycosylation sites of CFI. In some embodiments, the CFI variants comprise modifications at any one or more positions corresponding to positions N52, N85, N159, N446, N476, and N518 of a CFI having an amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI variants comprise one or more of substitutions selected from N52Q, N85Q, N159Q, N446Q, N476Q, and N518Q, where the positions correspond to positions of a CFI having an amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI variants comprise one or more of the following substitution combinations: N52Q, N85Q, N159Q, N446Q, N476Q, and N518Q, where the positions correspond to positions of a CFI having an amino acid sequence set forth in SEQ ID NO: 5.

[0026] In some embodiments, the CFI mutant comprises one or more modifications in the SPD domain of CFI, such as 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, or N420, having the amino acid sequence set forth in SEQ ID NO:5. Including modifications at any one or more positions corresponding to 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.

[0027] 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 positions 456 and 465 of CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0028] In some embodiments, the CFI mutants are 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, G406H, G406 I, G406K, G406L, G406M, G406N, G406P, G406Q, G406S, G406T, G406V, G406W, G406Y, Y408L, Y408F, Y4 08G, Y408P, Y408D, Y408A, Y408N, Y408T, Y408K, Y408R, Y408H, Y408I, Y408E, Y408M, Y408Q, Y408S , Y408W, Y408V, Q409G, E416A, K418G, N422K, D425A, D425K, D425R, D425G, R456A, R456N, E457G, E45 7A, E457D, E457F, E457H, E457I, E457K, E457L, E457M, E457N, E457P, E457Q, E457R, E457S, E457T, E 457W, E457Y, E457V, K458A, E461Q, E461K, E461R, E461H, E461G, E461A, E461D, E461F, E461I, E461LE461M, E461N, E461P, E461S, E461T, E461W, E461Y, E461V, R462K, R462A, R462D, F4 64Y, S465G, Q467K, Q467R, W468C, G469L, T495F, Y496L, D497E, S499G, I500K, A502S , K504Q, K504E, K504R, K504A, K504G, K504L, K504P, K504H, K504D, K504F, K504I, K 504M, K504N, K504S, K504T, K504V, K504W, K504Y, D506A, D506V, D506E, D506G, S507 and one or more substitutions selected from A, 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, the positions of which correspond to positions in a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0029] In some embodiments, the CFI mutants are selected from the group consisting of 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, and F53 7K, T495F, Y496L, D497E, S499G, I500K, G533A, K534Q, P535K, E536N, F537K, T495F, Y496L, D497E, S499G, I500K, G533A, K534Q, and P535K , E536N, F537K, Q467K, F537K, E530G, N531G, E530D, F537K, E457G, E461Q, E457G, E461G, Y408L, N531G, E457G, E461Q, N531G, E457G, E461 Q, I322V, V323I, I322V, V323I, R327P, A328C, W468C, A328C, W468C, K326Y, R327N, Y408L, N531G, E461Q, Y408L, N531G, E457G, E461Q, R4 62K, Y408L, N531G, E457G, E461Q, F464Y, Y408L, N531G, E457G, R462K, F464Y, Y408L, N531G, E461Q, R462K, F464Y, Y408L, E457G, E461Q , R462K, F464Y, E457G, N531G, E461Q, R462K, F464Y, Y408L, E457G, E461Q, R462K, N531G, E457G, E461Q, F464Y, E416A, D425R, Y408L, and N5 31G, E457G, E461Q, 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, K340 G, I345G, L364G, Y372G, W381G, V390G, W381G, P384A, V390G, W381G, P384G, V390G, N404G, Q409G, K418G, D425G, T346R, K504E, E530R, T346K, K504D, E530K, G344R, Y408L, N531G, G344K, Y408L, N53 1G, 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, E and one or more of the following combinations of substitutions selected from: 530K, N531G, Y408L, S507A, N531G, Y408L, N531G, E457G, E461Q, R462K, F464Y, S507A, E457G, S507A, and N531G, P535A, and S507A, the positions of which correspond to the positions of CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0030] In some embodiments, the CFI mutant comprises one or more modifications in the active site of CFI. In some embodiments, the CFI mutant comprises a modification at a position corresponding to position S507 of CFI having the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI mutant comprises a substitution S507A, which corresponds to position S507 of CFI having the amino acid sequence set forth in SEQ ID NO: 5.

[0031] In some embodiments, the CFI variant comprises an A chain and a B chain, and the CFI variant comprises an N-terminal to C-terminal structural arrangement of (A chain)-(optional linker)-(B chain). In some embodiments, the CFI variant comprises an A chain and a B chain, and the CFI variant comprises an N-terminal to C-terminal structural arrangement of (B chain)-(optional linker)-(A chain). In some embodiments, the CFI variant comprises a modification at one or more of C309 and C435, the positions of which correspond to those of a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0032] In some embodiments, the CFI mutant contains the substitutions C309S, C435S, which positions correspond to those of a CFI having the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the B and A chains are further linked by a disulfide bond.

[0033] In some embodiments, the CFI mutants are more prone to activation compared to wild-type CFI. In some embodiments, the CFI mutants contain modifications at any one or more positions corresponding to positions I317, R318, R319, K320, and R321 of a CFI having an amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the CFI mutants contain one or more substitutions selected from I317D, R318D, R319D, K320D, and R321K, where the positions correspond to positions of a CFI having an amino acid sequence set forth in SEQ ID NO: 5.

[0034] In some embodiments, the CFI mutant contains the substitutions I317D, R318D, R319D, K320D, and R321K, positions which correspond to positions in a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0035] In some embodiments, the CFI mutants are selected from the group consisting of 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, N5 31G, P535A, Y20F, N531G, P535A, E38A, N531G, P535A, S250A, N531G, P535A, D425A, N531G, P535A, Y20F, Y408L, N531G, E457G, E461Q, and R4 62K, F464Y, E38A, Y408L, N531G, E457G, E461Q, R462K, F464Y, S250A, Y408L, N531G, E457G, E461Q, R462K, F464Y, D425R, Y408L, and N531G , E457G, E461Q, R462K, F464Y, Y20F, E38A, S250A, D425A, Y20F, E38A, S250A, D425A, Y408L, N531G, E457G, E461Q, R462K, F464Y, Y20 F, 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, K5 04D, Y408L, N531G, K504E, Y408L, N531G, E457G, N531G, D425K, Y408F, N531G, Y408L, E457G, N531G, D425K, Y408L, E457G, P535G, D425K,The CFI comprises one or more of the following combinations of substitutions: Y408L, E457G, N531G, K534Q, Y408L, N531G, R462K, F464Y, and Y408L, P535G, and D425K, the positions of which correspond to the positions of CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0036] In some embodiments, the CFI variant comprises a CFI variant having an amino acid sequence as set forth in SEQ ID NO:5, which is comprised of a CFI variant having the amino acid sequence as set forth in SEQ ID NO:5, and a CFI variant having the amino acid sequence as set forth in SEQ ID NO:5, 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, N420, N422, N424, N426, N428, N429, N430, N431, N432, N433, N434, N435, N436, N437, N438, N439, N440, N441, N442, N443, N444, N445, N446, N447, N448, N449, N450, N451, N452, N453, N454, N455, N456, N457, N458, N459, N460, N461, N462, N463, N464, N465, N466, N467, N468, N469, N470, N471, N472, N473, N474, N475, N476, N477, N478, N479, N480, N481, N482, N483, 22, 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.

[0037] In some embodiments, the CFI mutants are K14A, K312A, R314A, I322T, I322Y, I322V, V323I, V323G, V323A, K326A, R327A, R327P, R327N, A328C, T377G, K340G, D341A, G344R, G344K, G344Y, I345G, T346R, T346K, T346H, A36 1G, L364G, Y372G, W381K, W381G, P384A, P384G, V390G, N402E, N404G, G406D, G406E, G406F, G406H, G4 06I, G406K, G406L, G406M, G406N, G406P, G406Q, G406S, G406T, G406V, G406W, G406Y, Y408L, Y408F, Y4 08G, Y408P, Y408D, Y408A, Y408N, Y408T, Y408K, Y408R, Y408H, Y408I, Y408E, Y408M, Y408Q, Y408S, Y 408W, Y408Y, Y408V, Q409G, E416A, K418G, N422K, D425A, D425K, D425R, D425G, R456A, R456N, E457G, E457A, 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, R462D, F464 Y, S465G, Q467K, Q467R, W468C, G469L, T495F, Y496L, D497E, S499G, I500K, A502S, K50 4Q, K504E, K504R, K504A, K504G, K504L, K504P, K504H, K504D, K504F, K504I, K504M, K 504N, K504S, K504T, K504V, K504W, K504Y, D506A, D506V, D506E, D506G, S507A, E530D, or consisting of any one or more substitutions selected from 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, the positions of which correspond to positions in a CFI having the amino acid sequence set forth in SEQ ID NO:5.

[0038] In some embodiments, the CFI mutants are 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+E457G+E 461Q-R462K+F464Y, Y408L+N531G+P535A, K14A+D425R, E530D+N531G+G533A+K534Q+P535K+E536N, A502S+K504Q+F537K, T495F+Y496L+D49 7E+S499G+I500K, G533A+K534Q+P535K+E536N+F537K, T495F+Y496L+D497E+S499G+I500K+G533A+K534Q+P535K+E536N+F537K, Q467K+F537 K. R327P, A328C+W468C, A328C+W468C+K326Y+R327N, Y408L+N531G+E461Q, Y408L+N531G+E457G+E461Q+R462K, Y408L+N531G+E457G+E461Q+F 464Y, Y408L+N531G+E457G+R462K+F464Y, Y408L+N531G+E461Q+R462K +F464Y, Y408L+E457G+E461Q+R462K+F464Y, E457G+N531G+E461Q+R46 2K+F464Y, Y408L+E457G+E461Q+R462K, N531G+E457G+E461Q+F464Y, E416A+D425R, Y408L+N531G+E457G+E461Q+R462K+F464Y+S507A, E457 G+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, K340G+I345G, L364G+Y372G, W381G+V390G, W381 G+P384A+V390G, W381G+P384G+V390G, N404G+Q409G, K418G+D425G, T346R+K504E+E530R, T34 6K+K504D+E530K, G344R+Y408L+N531G, G344K+Y408L+N531G, T346R+Y408L+N531G, T346K+Y4 08L+N531G, K504D+Y408L+N531G, K504E+Y408L+N531G, Y408L+E530R+N531G, Y408L+E530K+N 531G, 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, the positions of which correspond to positions of a CFI having the amino acid sequence set forth in SEQ ID NO: 5. For example, in some embodiments, a CFI variant comprises or consists of a substitution selected from N422K+E457G+N531G. For example, in some embodiments, a CFI variant comprises or consists of a substitution selected from T377G+E457G+N531G.

[0039] In some embodiments, the fusion construct comprises cfi and albumin, where albumin is human serum albumin (HSA) and CFI is wild-type CFI, such fusion constructs are referred to herein as "CFI-HSA". Exemplary CFI-HSAs of the present disclosure have an amino acid sequence of SEQ ID NO:21 or a sequence that comprises 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.

[0040] 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 comprises 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.

[0041] 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 comprises 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.

[0042] In some embodiments, the binding partner of CFI in the fusion construct comprises at least one domain or part of a domain of complement receptor 1 (CR1). In some embodiments, at least one CR1 domain comprises CCP domains 15-17 of CR1. In some embodiments, at least one CR1 domain comprises CCP domains 1-3 of CR1. In some embodiments, the fusion constructs of the present disclosure comprising at least one CR1 domain also comprise a fusion with albumin. In some embodiments, the fusion constructs of the present disclosure comprising at least one CR1 domain also comprise a fusion with albumin and / or at least one domain of factor H. In some embodiments, at least one CR1 domain comprises CR1 CCP domain 15. In some embodiments, at least one CR1 domain comprises CR1 CCP domain 16. In some embodiments, at least one CR1 domain comprises CR1 CCP domain 17. In some embodiments, at least one CR1 domain comprises CR1 CCP domains 15-16. In some embodiments, at least one CR1 domain comprises CR1 CCP domains 16-17. In some embodiments, an exemplary fusion construct comprises a CFI with the modification N531G fused to CCP domains 15-17 of CR1. In some embodiments, an exemplary fusion construct comprises a CFI fused to a CCP domain of CR1, further fused to albumin.

[0043] In some embodiments, the binding partner of CFI in the fusion construct comprises at least one domain or part of a domain of factor H. In some embodiments, at least one factor H domain comprises any one or more of Complement Control Protein (CCP) domains 1-20 of factor H. In some embodiments, the amino acid sequence of at least one factor H domain is or is derived from the sequence set forth in SEQ ID NO: 22 (Table 1). In some embodiments, at least one factor H domain comprises each of CCP domains 1-20 of factor H. In some embodiments, at least one factor H domain comprises CCP1, CCP2, CCP3, and CCP4. In some embodiments, at least one factor H domain comprises CCP2, CCP3, and CCP4. In some embodiments, at least one factor H domain comprises 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 set forth in SEQ ID NO: 23 (Table 1). In some embodiments, at least one factor H domain comprises factor H CCP domains 1-4 and 19-20. In some aspects, an exemplary fusion construct comprises CFI fused to a CCP domain of FH, further fused to albumin.

[0044] Exemplary CFI-HSA fusion constructs of the present disclosure include, represented from N-terminus to C-terminus: (a) N-terminal HSA-GGSSGG linker (SEQ ID NO:6)-CFI SEQ ID NO:5, containing substitutions N422K, E457G, and N531G. (b) N-terminal HSA-GGSSGG linker (SEQ ID NO:6)-CFI SEQ ID NO:5, containing substitutions T377G, E457G, and N531G.

[0045] An exemplary CR1-containing fusion construct of the present disclosure comprises HSA, CFI, and human CR1 domains. An example of such a fusion construct is the following: (a) Substitutions N422K, E457G, and N531G-GGSSGGSSGGSSGGSSGGSSGG linker (SEQ ID NO: 24)-N-terminal HSA-GGSSGG linker (SEQ ID NO: 6)-CFI SEQ ID NO: 5, including human CR1 domains CCP15, CCP16, and CCP17.

[0046] The components of the fusion construct of the present disclosure (CFI and binding partner) may be held together by an optional linker. They may be of any suitable length of at least one amino acid. The linker may be a flexible linker and may be a peptide of about 1 to about 20 amino acid residues in length, and the amino acid residues may include glycine residues. The linker may also optionally include serine residues. 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 linkers known in the art. An exemplary linker is (GGSS) n GG (SEQ ID NO: 25), where n is any number from about 1 to about 25. In some embodiments, the linker is a protease-sensitive cleavable linker. Exemplary linkers linking 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.

[0047] In some embodiments, CFI-HSA may have an extended half-life relative to a CFI that is not part of a fusion construct. An exemplary CFI-HSA construct may be generated by linking albumin to wild-type CFI by 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.

[0048] In some embodiments, the fusion construct comprises an N-terminal to C-terminal structural arrangement such as (albumin)-(optional linker)-(WT CFI).

[0049] In some embodiments, the fusion construct comprises an N-terminal to C-terminal structural arrangement such as (WT CFI)-(optional linker)-(albumin).

[0050] 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, and the fusion construct comprises the structural arrangement N-terminal to C-terminal (SEQ ID NO:7)-(optional linker)-(SEQ ID NO:5). In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:5 and SEQ ID NO:7, and the fusion construct comprises the structural arrangement N-terminal to C-terminal (SEQ ID NO:7)-(linker)-(SEQ ID NO:5). 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, and the fusion construct comprises the structural arrangement N-terminal to C-terminal (SEQ ID NO:7)-(SEQ ID NO:6)-(SEQ ID NO:5). In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:5 and SEQ ID NO:7, and the fusion construct comprises the structural arrangement N-terminal to C-terminal (SEQ ID NO:5)-(optional linker)-(SEQ ID NO:7). In some embodiments, the fusion construct comprises the amino acid sequences of SEQ ID NO:5 and SEQ ID NO:7, and the fusion construct comprises the structural arrangement from N-terminus to C-terminus: (SEQ ID NO:5)-(linker)-(SEQ ID NO:7). 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, and the fusion construct comprises the structural arrangement from N-terminus to C-terminus: (SEQ ID NO:5)-(SEQ ID NO:6)-(SEQ ID NO:7).

[0051] In some embodiments, the fusion construct comprises an amino acid sequence set forth in SEQ ID NO:21, or an amino acid sequence comprising 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 set forth 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. It is noted that in some embodiments, the albumin fusion to wild-type CFI (e.g., N-terminal albumin fusion) provides solubility and facilitates activation of CFI-HSA. When activation of CFI to a mature two-chain protein by Furin is performed post-translationally and activation is compared between CFI-HSA and wild-type CFI without albumin (WT-CFI), it is observed that Furin activates CFI-HSA significantly better and almost completely. It is observed that the CFI-HSA protein remains monomeric and there is no evidence of aggregation. There are significant and unexpected benefits of the amino-terminal HSA fusion to maintain the solubility, monodispersity, and efficient Furin activation of the CFI-HSA construct, including significantly improved bioavailability through improved half-life.

[0052] Thus, provided herein is a method for increasing the activation of CFI, comprising fusing HSA to wild-type CFI, the fusion being an N-terminal fusion prior to activation by furin, and activating by furin. In some embodiments, activation by furin is performed in cells during recombinant production of the CFI fusion construct of the present disclosure. In some embodiments, activation by furin is performed in vitro.

[0053] With reference to Table 1, SEQ ID NO:1 is the amino acid sequence of wild-type plasma-derived human CFI, designated "CFI-PD," and has a leader sequence. The wild-type CFI used in fusion with a second component may comprise the amino acid sequence of SEQ ID NO:5, without the leader sequence present in SEQ ID NO:1. Alternatively, the mouse Ig kappa chain V-III region MOPC63 leader sequence (SEQ ID NO:2) may be used in recombinant production of any of the CFI fusion constructs provided herein. In some embodiments, provided herein are CFI fusion constructs 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.

[0054] Table 1: Components of exemplary CFI fusion constructs [Table 1-1]

[0055] (Continuation of Table 1) [Table 1-2]

[0056] II. Generation of CFI fusion constructs Cell lines can be developed to express the production of CFI and fusion constructs described herein. Cell lines for producing fusion constructs can be produced using any host cell capable of expressing the fusion constructs described herein. The host cells can be mammalian, insect, fungal, plant, and / or bacterial cells. For expression of CFI fusion constructs, 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 is a mammalian cell line. In some embodiments, the host cell is a Chinese Hamster Ovary (CHO) cell.

[0057] The CFI fusion constructs described herein can be recombinantly expressed in mammalian cell lines known in the art for producing biological products, 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.

[0058] Methods for the production and purification of the CFI fusion constructs described herein are provided herein. The CFI fusion constructs described herein may be purified from conditioned media by standard methods known in the art. In some embodiments, the CFI fusion constructs may be purified by chromatography on an affinity matrix. In some embodiments, the affinity matrix is ​​CaptureSelect™ human albumin affinity matrix. In some embodiments, the CFI fusion constructs may be purified by chromatography on a cation exchange matrix and / or anion exchange matrix, and optionally size exclusion chromatography. The CFI fusion constructs may be optimally buffer exchanged into any suitable buffer known in the art. Purity may be assessed by any method known in the art, including gel electrophoresis, orthogonal HPLC methods, staining and spectrophotometric techniques.

[0059] III.CFI Fusion Construct - How to Use A.Use The CFI fusion constructs of the present disclosure may be used to regulate the complement system in the eye.

[0060] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of regulating the classical and lectin complement pathways in the eye.

[0061] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of regulating the alternative complement pathway.

[0062] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing the amplification of the complement system in the eye.

[0063] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing cleavage of C3b in the eye.

[0064] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing cleavage of C4b in the eye.

[0065] 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.

[0066] 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.

[0067] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the generation of C3dg from iC3b in the eye.

[0068] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the generation of C3c from iC3b in the eye.

[0069] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of reducing levels of C3bα chain in the eye.

[0070] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing hydrolysis of peptide substrates in the eye.

[0071] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of increasing the proteolysis of macromolecular protein substrates in the eye.

[0072] 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.

[0073] As discussed herein, in some embodiments, the CFI fusion constructs of the present disclosure are capable of modulating the hemolysis observed in the eye.

[0074] 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.

[0075] 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.

[0076] The CFI fusion constructs of the present disclosure may be used to treat the eye of a subject. As used herein, a subject includes any mammalian subject, including 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.

[0077] B. Treatment of Ocular Symptoms The CFI fusion constructs provided herein are useful for treating an ocular condition in a subject. In some embodiments, methods are provided for treating an ocular condition in a subject in need thereof, comprising intraocularly administering to the subject a therapeutically effective amount of any of the CFI fusion constructs provided herein, or a pharmaceutical composition thereof, at a dose of about 0.1 to about 10 mg per eye and / or at a frequency of about every 1 to about 4 months.

[0078] In some embodiments, the ocular condition is characterized by a deficiency in CFI, hi some embodiments, the ocular condition is characterized by dysregulation of the complement system.

[0079] In some embodiments, the ocular condition is characterized by the presence of a dysfunctional CFI gene, hi some embodiments, the ocular condition is characterized by dysregulation of the complement system and low CFI levels.

[0080] In some embodiments, the ocular condition 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 vitreous retinopathy, and age-related macular degeneration (AMD).

[0081] In some embodiments, the ocular condition is dry AMD. In some embodiments, the ocular condition is early stage dry AMD. In some embodiments, the ocular condition is intermediate stage dry AMD. In some embodiments, the ocular condition is late stage dry AMD. In some embodiments, the ocular condition is late stage dry AMD that has progressed to wet AMD. In some embodiments, the ocular condition is late stage dry AMD that has progressed to geographic atrophy (GA).

[0082] In some embodiments, the ocular condition is wet AMD. In some embodiments, the ocular condition is early stage wet AMD. In some embodiments, the ocular condition is intermediate stage wet AMD. In some embodiments, the ocular condition is late stage wet AMD. In some embodiments, the ocular condition is late stage wet AMD that has progressed to neovascular AMD.

[0083] In some embodiments, the ocular condition is geographic atrophy (GA). In some embodiments, the ocular condition is GA that has progressed from dry AMD. In some embodiments, the GA is associated with late-stage AMD. In some embodiments, the GA is not associated with late-stage AMD.

[0084] In some embodiments, the ocular condition is caused by one or more mutations to the CFI gene in 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.

[0085] In some embodiments, the ocular condition is Phe13Val, Val20Ile, His40Arg, Cys43Phe, Asp44Asn, Pro50Ala, Cys54Arg, Cys54 * , Ile55Phe, Pro64Leu, Asn70Thr, Thr72Ser, Phe82Ser, Pro83Gln, Ser90Asn, Cys106Arg, Thr107Ala, Glu109 Ala, His118Arg, Gly119Arg, Gly125Arg, Val127Ala, Val129Gly, Leu131Arg, Met138Val, Met138Ile, Trp145 * , Asn151Ser, Val152Met, Gly162Asp, Asp164Val, Asn177Ile, His183Arg, Val184Met, Arg187 *, Arg187Gln, Gly188Ala, Phe198Leu, Arg202Ile, Thr203Ile, Tyr206Asn, Gln217His, Ser221Tyr, Asp224Asn, Cys229Arg, Val230Met, Val230Glu, Ala240Gly, Cy s247Gly, Gly248Glu, Asp249Glu, Ala258Thr, Gly261Ser, Gly261Asp, Gly263Val, Lys267Asn, Gly280Asp, Gly287Arg, Glu290Asp, Ala300Thr, Glu303Lys, Glu305 * , Ile306Val, Ile306Ser, Asp310Glu, Arg317Trp, Arg317Gln, Cys327Arg, Gly328Arg, Arg339 * , Arg339Gln, Ile340Thr, Gly342Glu, Arg345Gln, Gly349Arg, Val355Met, Ala356Pro, Ile357Met, Gly362A la, Tyr369Ser, Trp374Cys, Arg389His, Trp399Arg, Pro402Ser, Asp403Asn, Arg406Cys, Arg406His, Tyr411 * , Val412Met, Ile416Leu, His418Leu, Gly424Asp, Asp429Glu, Ala431Thr, Ile433Val, Ile433T hr, 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 *The CFI gene is caused by one or more mutations in the subject selected from the group consisting of:

[0086] In some embodiments, the ocular condition is caused by one or more of a group of mutations in a subject with GA consisting of C2, CFB, CFH, and C3 mutations.

[0087] C. Combined therapy Administration of any one of the therapeutic CFI fusion constructs provided herein may be a monotherapy or may be in combination with any other known drug or treatment for a condition associated with dysregulation of the complement system or associated with a CFI deficiency.

[0088] D. Administration The CFI fusion constructs described herein are delivered to the eye as a polypeptide-based therapy.

[0089] Such treatments contemplated herein include administration of a CFI fusion construct of the present disclosure to the eye. Accordingly, pharmaceutical compositions comprising a CFI fusion construct of the present disclosure are provided herein.

[0090] Ocular administration of the therapeutic CFI fusion constructs described herein may be intraorbital, periocular, intraocular, intravitreal, or subretinal. Administration of the fusion construct may be performed using any suitable excipient, carrier, or other agent to provide suitable or improved tolerance, mobility, delivery, etc. In an exemplary embodiment, administration of the CFI fusion constructs described herein is intravitreal.

[0091] In some embodiments, the CFI fusion constructs are expressed as inactive single-chain precursor proteins and can be activated by furin, a serine protease. Furin is an endoprotease that cleaves CFI at its conserved RRKR sequence (also referred to as the furin recognition sequence), resulting in heavy and light chains linked by disulfide bonds. The furin-processed mature two-chain protein is the activated form of the CFI protein. As generally contemplated herein, the CFI fusion constructs described herein are delivered in an activated two-chain form. However, in some cases, the inactive CFI fusion constructs can be delivered in an inactive single-chain form. In some embodiments, the delivered form includes both the single-chain inactive form and the two-chain active form.

[0092] E. Medication In embodiments provided herein, the CFI fusion constructs described herein are administered to one or both eyes of a subject in need thereof at a dose of about 0.1 mg to about 10 mg / eye and / or at a frequency of about every 4 to 16 weeks.

[0093] The optimal dose ranges and frequencies of the present disclosure are expected to maintain baseline CFI levels above those in subjects afflicted with an ocular disease (e.g., AMD) for 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 the CFI fusion construct.

[0094] In some embodiments, upon ocular administration of the fusion CFI construct, the baseline CFI level of a subject suffering from an ocular disease is maintained above the baseline CFI level of a reference subject suffering from the same ocular disease who did not receive the injection, hi some embodiments, such CFI level is maintained for at least 10, 15, 2025, 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 injection of CFI-HSA.

[0095] 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 to about 10 mg / eye at a frequency of about every 4 to 16 weeks.

[0096] 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 two eyes sequentially.

[0097] Exemplary dosages of the CFI fusion constructs described herein are from about 0.1 mg to about 10 mg / eye. In some embodiments, the dosage of a CFI fusion construct described herein is about 0.5 mg to about 10 mg / eye, about 1 mg to about 10 mg / eye, about 1.5 mg to about 10 mg / eye, about 2 mg to about 10 mg / eye, about 2.5 mg to about 10 mg / eye, about 3 mg to about 10 mg / eye, about 3.5 mg to about 10 mg / eye, about 4 mg to about 10 mg / eye, about 4.5 mg to about 10 mg / eye, about 5 mg to about 10 mg / eye, about 5.5 mg to about 10 mg / eye, about 6 mg to about 10 mg / eye, about 6.5 mg to about 10 mg / eye, about 7 mg to about 10 mg / eye, about 7.5 mg to about 10 mg / eye, about 8 mg to about 10 mg / eye, about 8.5 mg to about 10 mg / eye, about 9 mg to about 10 mg / eye, or about 9.5 mg to about 10 mg / eye.

[0098] In some embodiments, administration of the CFI fusion constructs described herein is intraocular (e.g., intravitreal) administration at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye.

[0099] 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 frequency of administration of the CFI fusion construct is intravitreal 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.

[0100] In some embodiments, administration of a therapeutic CFI fusion construct described herein is intraocular (e.g., intravitreal) administration at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 4 weeks.

[0101] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 5 weeks.

[0102] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 6 weeks.

[0103] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 7 weeks.

[0104] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 8 weeks.

[0105] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 9 weeks.

[0106] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 10 weeks.

[0107] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 11 weeks.

[0108] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 12 weeks.

[0109] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 13 weeks.

[0110] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 14 weeks.

[0111] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 15 weeks.

[0112] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / eye, about 0.5 mg / eye, about 1 mg / eye, about 1.5 mg / eye, about 2 mg / eye, about 2.5 mg / eye, about 3 mg / eye, about 3.5 mg / eye, about 4 mg / eye, about 4.5 mg / eye, about 5 mg / eye, about 5.5 mg / eye, about 6 mg / eye, about 6.5 mg / eye, about 7 mg / eye, about 7.5 mg / eye, about 8 mg / eye, about 8.5 mg / eye, about 9 mg / eye, about 9.5 mg / eye, or about 10 mg / eye at a frequency of about every 16 weeks.

[0113] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.1 mg / 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.

[0114] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 0.5 mg / 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.

[0115] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 1.0 mg / 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.

[0116] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 1.5 mg / 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.

[0117] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 2 mg / 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.

[0118] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 2.5 mg / 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.

[0119] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 3 mg / 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.

[0120] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 3.5 mg / 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.

[0121] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 4 mg / 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.

[0122] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 4.5 mg / 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.

[0123] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 5 mg / 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.

[0124] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 5.5 mg / 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.

[0125] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 6 mg / 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.

[0126] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 6.5 mg / 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.

[0127] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 7 mg / 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.

[0128] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 7.5 mg / 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.

[0129] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 8 mg / 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.

[0130] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 8.5 mg / 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.

[0131] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 9 mg / 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.

[0132] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 9.5 mg / 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.

[0133] In some embodiments, administration of the CFI fusion construct is intraocular (e.g., intravitreal) at a dose of about 10 mg / 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.

[0134] F. Preparation Pharmaceutical compositions containing the CFI fusion constructs of the present disclosure can be formulated in any conventional manner suitable for ocular administration by mixing an amount of the polypeptide selected for use in the treatments provided herein with one or more physiologically acceptable carriers or excipients. The choice of carrier or excipient is within the skill of the administration specialist and can depend on several parameters. These include, for example, the mode of administration and the disorder being treated. The pharmaceutical compositions provided herein can be formulated for single-dose (direct) administration or for dilution or other modification. The concentration of the compound in the formulation is effective for delivery of an amount effective for the intended treatment upon administration. Typically, but not necessarily, the composition is formulated for single administration.

[0135] In some exemplary embodiments, the fusion construct may be concentrated to at least 150 mg / mL. In some embodiments, the fusion construct may be dosed up to at least 7.5 mg in 50 uL. In some embodiments, the fusion construct may be dosed up 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, about 90 uL to about 150 uL, about 100 uL to about 150 uL, about 120 uL to about 150 uL, about 140 uL to about 150 uL, about 160 uL to about 150 uL, about 180 uL to about 150 uL, about 190 uL to about 200 uL, about 210 uL to about 250 uL, about 220 uL to about 250 uL, about 230 uL to about 250 uL, about 240 uL to about 250 uL, about 250 uL to about 250 uL, about 260 uL to It may be administered so that at least 7.5 mg is administered in 0 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.

[0136] In some exemplary embodiments, the fusion construct is at least 7.5 mg in 25 uL, at least 7.5 mg in 30 uL, at least 7.5 mg in 35 uL, at least 7.5 mg in 40 uL, at least 7.5 mg in 45 uL, at least 7.5 mg in 50 uL, at least 7.5 mg in 55 uL, at least 7.5 mg in 60 uL, at least 7.5 mg in 65 uL, at least 7.5 mg in 70 uL, at least 7.5 mg in 75 uL, at least 7.5 mg in 80 uL, at least 7.5 mg in 85 uL, at least 7.5 mg in 90 uL, at least 7.5 mg in 100 uL, at least 7.5 mg in 120 uL, at least 7.5 mg in 140 uL, at least 7.5 mg in 160 uL, at least 7.5 mg in 180 uL, at least 7.5 mg in 190 uL, at least 7.5 mg in 200 uL, at least 7.5 mg in 210 uL, at least 7.5 mg in 220 uL, at least 7.5 mg in 230 uL, at least 7.5 mg in 240 uL, at least 7.5 mg in 250 uL, at least 7.5 mg in 260 uL, at least 7.5 mg in 280 uL, at least 7.5 mg in 290 uL, at least 7.5 mg in 300 uL, at least 7.5 mg in 310 uL, at least 7.5 mg in 320 uL, at least The dosage may be at least 7.5mg in 0 uL, at least 7.5mg in 95 uL, at least 7.5mg in 100 uL, at least 7.5mg in 105 uL, at least 7.5mg in 110 uL, at least 7.5mg in 115 uL, at least 7.5mg in 120 uL, at least 7.5mg in 125 uL, at least 7.5mg in 130 uL, at least 7.5mg in 135 uL, at least 7.5mg in 140 uL, at least 7.5mg in 145 uL, or at least 7.5mg in 150 uL.

[0137] In some exemplary embodiments, the fusion construct is dosed to provide about 5 mg to 10 mg in 25 uL, 30 uL, 35 uL, 40 uL, 45 uL, 50 uL, 55 uL, 60 uL, 65 uL, 70 uL, 75 uL, 80 uL, 85 uL, 90 uL, 95 uL, 100 uL, 105 uL, 110 uL, 115 uL, 120 uL, 125 uL, 130 uL, 135 uL, 140 uL, 145 uL, or 150 uL.

[0138] G. Pharmaceutical Compositions The present disclosure also provides a pharmaceutical composition comprising any one of the CFI fusion constructs disclosed herein and, optionally, a pharma- ceutically acceptable excipient or carrier suitable for ocular administration. In some embodiments, the pharmaceutical composition is sterile. The pharmaceutical composition may be formulated to be compatible with their 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.

[0139] H. Kits and Articles of Manufacture for Therapeutic CFI Fusion Constructs The present disclosure also provides a kit or article of manufacture comprising any one of the CFI fusion constructs disclosed herein or any pharmaceutical composition disclosed herein. In some embodiments, the kit may further comprise instructional materials for carrying out any of the dosing and intraocular administration methods disclosed herein. In some embodiments, the kit may further comprise a sterile container or vial for holding the fusion construct and / or pharmaceutical composition disclosed herein. In some embodiments, the kit may further comprise a sterile delivery device for administering the fusion construct and / or pharmaceutical composition disclosed herein. In some embodiments, the article of manufacture comprises any pharmaceutical composition of the present disclosure. EXAMPLES

[0140] Example 1: Expression, purification, activation, and in vitro sialylation of CFI-HSA overview In Example 1, reference to CFI-HSA refers to human serum albumin fused to the N-terminus of wild-type CFI (SEQ ID NO:21).

[0141] Wild-type CFI-HSA protein was expressed in Chinese Hamster Ovary (CHO) cells, purified by anti-albumin affinity purification, activated with Furin, and purified by sizing column. The activated CFI-HSA protein was subjected to in vitro sialylation to increase the total sialylation of CFI-HSA. Finally, the sialylated protein was purified using anti-albumin affinity purification and finalized by size-exclusion column chromatography.

[0142] Expression The CFI-HSA gene (SEQ ID NO: 21) with human serum albumin at the amino terminus of the CFI protein was synthesized (ThermoFisher Scientific, Geneart, Regensburg, Germany). The protein was made with a signal sequence of SEQ ID NO: 2 that 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 (Lake Pharma, Hayward, CA) using standard molecular biology techniques. The resulting plasmid DNA was transformed into E. coli. The transfected E. coli was grown in 200 mL LB medium for expression of the plasmid DNA and harvested using standard techniques. The plasmid DNA was run on an agarose gel for quality assessment and sequence confirmation before proceeding to transfection.

[0143] 1.0 liter of suspension TunaCHO™ cells were seeded into shake flasks and grown using serum-free synthetic medium. On the day of transfection, the grown cells were seeded into new flasks containing fresh medium. Plasmid DNA was transiently transfected into CHO cells using Lipofectamine 2000 (ThermoFisher Scientific). Cells were maintained as batch-fed cultures until the end of the production run. Protein was expressed for 14 days at 37°C, 125 RMP with 8% CO2 concentration. Cells were centrifuged and supernatant was collected for purification of secreted CFI-HSA after 14 days of expression.

[0144] purification The supernatant containing the expressed CFI-HSA protein was passed through a 10 mL gravity-flow column of CaptureSelect™ human albumin affinity matrix (ThermoFisher Scientific). The protein bound to the column 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 containing 2 M MgCl2, and second, with 3 column volumes of 20 mM citric acid, pH 3.0. The eluates from both step 1 and step 2 were collected in 5 mL fractions. Each fraction of the step 2 elution was neutralized with 10% neutralization buffer (1.5 M tris-HCL pH 7.4). All fractions were analyzed by reducing and non-reducing SDS-PAGE electrophoresis, and bands were visualized by SimplyBlue™ SafeStain (ThermoFisher Scientific). CFI-HSA migrates as a 130 kDa band on non-reducing gels and as 102 kDa and 28 kDa bands on reducing gels. Fractions with the greatest CFI-HSA concentration and purity were pooled for further processing.

[0145] Furin activation Cleavage of CFI-HSA to produce the protein in its activated form was performed by incubating 4 μg of recombinant Furin per mg of purified CFI-HSA in Tris-NaCl, 2.5 mM CaCl2, and 0.5% CHAPS for 18 h at 30°C. The CFI-HSA protein concentration was maintained at 1.4 mg / mL, which results in more than 90% activation of the protein. The activated protein was separated from inactivated CFI-HSA and other proteins by size exclusion chromatography. Size exclusion chromatography (SEC) 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 Na2HPO4, 2 mM KH2PO4, pH 7.4) as the mobile phase. The collected fractions were analyzed by CE-SDS (LabChip GXII, Perkin Elmer). Fractions containing the target protein were pooled and analyzed by SE-UPLC.

[0146] In vitro sialylation The activated CFI-HSA protein was subjected to in vitro sialylation. Briefly, sialylation was performed in a two-step enzymatic reaction. First, the galactosylation reaction of CFI-HSA was carried out in a volume of 200 μL utilizing a 1:200 molar ratio of galactosyltransferase (GalT1) enzyme and CFI-HSA in 10 mM UDP-galactose, 5 mM MnCl2, and 100 mM MES, pH 6.5 buffer. Galactosylated CFI-HSA was purified from the reaction mixture by CaptureSelect™ human albumin affinity chromatography as previously described. The sialylation reaction was then carried out in a 250 μL volume at 37° C. for 1 h using a 1:50 molar ratio of the enzyme α 2,6-sialyltransferase and purified CFI-HSA in 80 μM alkaline phosphatase, 6.1 mM CMP-NANA, 10 mM ZnCl2, and 200 mM MES buffer, pH 6.5. Sialylated CFI-HSA protein was purified from the reaction mixture by CaptureSelect™ human albumin affinity chromatography. The extent and characteristics of sialic acid chains on CFI-HSA were determined by utilizing Agilent / Prozyme analytical services, the GS-SAP method for total sialic acid quantification (Agilent GS48), and mass spectrophotometric (MS) analysis (Lake Pharma analytical services), as described in more detail below.

[0147] Briefly, total sialic acid quantification was performed by mixing 20 μL of each sample with 10 μL of releasing reagent in a 96-well plate. The reaction mixture was incubated at 80° C. for 2 h. The samples were cooled to room temperature and 10 μL of labeling reagent was added to each sample for further incubation at 50° C. for 3 h. The samples were cooled again to room temperature and 160 μL of deionized (dI) water was added to a total volume of 200 μL. 10 μL of sample was injected onto an Agilent UHPLC Poroshell C18 column and run at 30° C. in 4% methanol, 8% acetonitrile in water (line A1) and 100% ACN (line B1) at a flow rate of 0.4 mL / min. Peaks were recorded at 373 / 448 nm wavelength. A standard curve of total peak area versus picomoles (pmol) of sialic acid was generated by running 1-2000 pmol of NANA (N-acetylneuraminic acid, Neu5Ac) supplied with the kit on the same column. Total sialic acid was quantified for each sample by comparing the peak area of ​​the sample to the standard curve. The resulting sialylation is summarized in Table 1.1 below.

[0148] Table 1.1: Sialylation assay results [Table 2]

[0149] Mass spectrometry was performed by a standard trypsin Q-TOF mass spectrometer. Briefly, all samples were treated with DTT and iodoacetamide, reduced, alkylated, and subsequently digested with trypsin. 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.

[0150] Table 1.2: Peptide analysis results [Table 3]

[0151] Finishing 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 pooled, the concentration was brought to 5 mg / mL, and the samples were flash-frozen for storage at -80°C.

[0152] N-terminal albumin fusion provides solubility and promotes activation of CFI-HSA Activation was compared between CFI-HSA and wild-type CFI (WT-CFI) without albumin or other fusion tags. Gene constructs for WT-CFI were expressed essentially as described above for CFI-HSA. Recombinant WT-CFI protein showed moderate purity by reducing SDS-PAGE, but significant High Molecular Weight Species (HMWS) and aggregates were observed under reducing and non-reducing conditions. For CFI-HSA with the addition of an N-terminal HSA tag, transient expression using TunaCHO™ cells followed by purification as described above showed no HMWS or aggregates on reducing and non-reducing SDS-PAGE. Activation of purified recombinant CFI with Furin resulted in a further increase in aggregates and HMWS with almost complete polydispersity. Furthermore, essentially no activated CFI was observed on reducing SDS-PAGE. In contrast, addition of Furin efficiently activated CFI-HSA almost completely under the same conditions, and the CFI-HSA protein remained monomeric under non-reducing conditions with no evidence of aggregates or HMWS. There are significant and unexpected benefits of the N-terminal HSA tag for maintaining solubility, monodispersity, and efficient Furin activation when compared to CFI lacking any fusion tag.

[0153] Example 2: Half-life of CFI-HSA in the vitreous humor of non-human primates In Example 2, references to CFI-HSA refer to human serum albumin fused to the N-terminus of wild-type CFI (SEQ ID NO:21).

[0154] Pharmacokinetics and pharmacodynamics of CFI-HSA after intravitreal injection The ocular pharmacokinetics of an N-terminal albumin fusion of wild-type CFI-HSA was investigated after intravitreal administration to six AGMs (African Green Monkeys). Six animals were divided into two groups 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 PBS for injection as a vehicle control. Non-terminal vitreous fluid samples (100 μL) were collected on days 1, 7, 14, 21, and 28 after administration. Vitreous fluid CFI-HSA drug concentrations were determined using a quantitative electrochemiluminescence (ECL) antigen assay optimized to measure CFI-HSA in the vitreous fluid of AGMs. The assay captures CFI-HSA levels using coating of 2 μg / mL of anti-CFI antibody (clone OX21, LifeSpan Biosciences (LS Bio), Seattle WA) onto assay plates from Meso Scale Discovery (MSD, Rockville, MA). Detection of captured CFI-HSA is performed using 0.5 μg / mL of goat polyclonal anti-HSA antibody (Abcam, Cambridge, MA) conjugated to SULFO-TAG, which emits light upon application of an electrical potential [electrochemiluminescence (ECL)]. ECL Relative Light Units (RLU) are measured with a MESO™ SECTOR S 600 reader and unknown CFI-HSA concentrations in the vitreous humor are interpolated from a standard curve ranging from 0.05 μg / mL to 40 μg / mL of Factor I-HSA. Data are provided in Table 2.1.

[0155] Table 2.1 CFI-HSA levels 1, 7, 14, 21, and 28 days after intravitreal administration [Table 4] * BLQ: Measurement below the limit of quantitation of the assay

[0156] Non-compartmental analysis yielded apparent terminal ocular half-lives of 3.6 and 4.1 days for the 250 μg and 500 μg dose levels, respectively.

[0157] Table 2.2. Estimated PK parameters for CFI-HSA in African green monkey vitreous humor [Table 5]

[0158] Complement component 3a (C3a) levels in the vitreous humor were determined by ELISA using the Quidel kit for C3a ELISA for doses of 250 μg and 500 μg ( FIG. 2 ). CFI-HSA fusion constructs dose-dependently reduced ocular C3a levels up to 7 days after intraocular injection. Increased C3b degradation by CFI-HSA reduces complex formation between C3b and Bb, leading to reduced C3 cleavage into C3a and C3b via the amplification loop of the alternative pathway.

[0159] The ocular pharmacokinetics of an N-terminal albumin fusion of wild-type CFI-HSA was investigated after intravitreal administration to six AGMs. Six animals were divided into two groups 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 a control group received a single intravitreal injection of vehicle (OU, N=4). The eyes (OU) of all 12 animals were injected with equal volumes of 100 μL of test article or vehicle control. Animals underwent ocular examinations (see Example 4) on days 0, 1, 7, or 8, 14, or 15, 21, and 28 after administration. Animals were euthanized on days 1, 7, 14, and 28 for PK-PD analysis and histopathology. Vitreous CFI-HSA drug concentrations were determined using a quantitative electrochemiluminescence (ECL) antigen assay optimized for measuring CFI-HSA in the vitreous humor of AGMs, as described above. Vitreous terminal half-life measurements were assessed to be 4 and 4.4 days for the 0.25 mg and 1.0 mg doses, respectively (Figure 3A).

[0160] Complement component 3b (C3b) and cleavage fragment iC3b levels in the vitreous humor were determined by chemiluminescence Western blot using anti-C3 antibody and quantified by densitometry for the 1.0 mg dose (FIG. 2B). The CFI-HSA fusion construct reduced ocular iC3b levels up to 14 days after intraocular injection. The increase in C3b degradation by CFI-HSA is expressed as the ratio of C3b / iC3b for days 1, 7, and 14, showing a significant increase in cleavage in eyes receiving 1.0 mg CFI-HSA injections compared to vehicle controls (FIG. 2B).

[0161] Example 3: Pharmacokinetic Modeling of CFI-HSA Intravitreal Injection for Human Dosing The half-life of CFI-HSA following intravitreal injection was measured over time in non-human primates. A half-life of approximately 4 days was found, preserving dose proportionality across the three dose levels in the AGM, as illustrated in Figure 3A. Furthermore, CFI-HSA cleavage activity was detected via Western blot for up to 14 days (Figure 2B).

[0162] Based on the AGM data, a vitreous PK model for human dosing was generated, as illustrated in FIG. 3B. Without being bound by theory or mechanism, intravitreal injection of CFI-HSA at 1 mg / eye is predicted to maintain a baseline CFI level above that of AMD patients for 60 days after injection. Intravitreal injection of CFI-HSA at 10 mg / eye is predicted to maintain a baseline CFI level above that of AMD patients for 90 days after injection.

[0163] Example 4: Safety Profile of CFI-HSA Intravitreal Injection in Non-Human Primates The safety profile of intravitreal injection of CFI-HSA was determined in non-human primates. After injection, AGMs were evaluated for clinical scores via slit lamp examination on days 0, 1, 7, or 8, 14, or 15, 21, and 28, or until termination. As illustrated in FIG. 4A, clinical scores using slit lamp examination were made over time after intravitreal injection of CFI-HSA. AGM subjects were also evaluated for intraocular pressure after injection of CFI-HSA. As illustrated in FIG. 4B, intraocular pressure of injected AGM subjects was measured over time. The intraocular pressure of each injected AGM subject was found to be within baseline fluctuations and therefore stable. Finally, retinal thickness of AGM subjects was evaluated after injection of CFI-HSA. As illustrated in FIG. 4C, retinal thickness of injected AGM subjects was measured over time. The retinal thickness of each injected AGM subject was found to be within baseline variation and therefore stable.

Claims

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

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

5.

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

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

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

6. 2. The method of claim 1, wherein the fusion protein comprises complement factor I (CFI) comprising 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 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof.

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

7.

9. The method of any one of claims 1 to 8, wherein the fusion construct comprises 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 of any one of claims 1 to 9, wherein the fusion construct comprises the amino acid sequence of SEQ ID NO:

21.

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

12. 12. The method of claim 11, wherein the binding partner comprises a cofactor of factor H 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. 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. 15. The method of claim 14, wherein the variant of serum albumin comprises at least 70%, at least 80%, at least 90%, or at least 95% sequence identity to 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 eye condition is geographic atrophy (GA).

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

20. 20. The method of any one of claims 1 to 19, wherein the human subject carries one or more mutations to 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 dose may be from about 0.1 mg to about 10 mg / eye, from about 0.5 mg to about 10 mg / eye, from about 1 mg to about 10 mg / eye, from about 1.5 mg to about 10 mg / eye, from about 2 mg to about 10 mg / eye, from about 2.5 mg to about 10 mg / eye, from about 3 mg to about 10 mg / eye, from about 3.5 mg to about 10 mg / eye, from about 4 mg to about 10 mg / eye, from about 4.5 mg to about 10 mg / eye, from about 5 mg to about 10 mg / eye, 21. The method of any one of claims 1 to 20, wherein the dose is about 5.5 mg to about 10 mg / eye, about 6 mg to about 10 mg / eye, about 6.5 mg to about 10 mg / eye, about 7 mg to about 10 mg / eye, about 7.5 mg to about 10 mg / eye, about 8 mg to about 10 mg / eye, about 8.5 mg to about 10 mg / eye, about 9 mg to about 10 mg / eye, or about 9.5 mg to about 10 mg / eye.

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

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

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

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

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

32. 31. The method of any one of claims 1 to 30, wherein upon administration, the baseline CFI level is maintained above that of a reference subject who did not receive the injection.

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