Methods for treating complement-mediated diseases

Abstract

Provided herein are methods of treating condensation set disease (CAD) or chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof. These methods comprise administering to a subject a humanized antibody (anti-C1s antibody) that specifically binds to the complement component C1s. A method of treating CAD comprises administering the anti-C1s antibody to a subject at a fixed dose. In some embodiments, methods of treating CIDP comprise administering to a subject a weight-based load dose of the anti-C1s antibody and then administering one or more fixed maintenance doses. These methods include administering an effective dose of an anti-C1s antibody to achieve a minimum level of CP inhibition for therapeutic effects.

Description

[0001] Related Applications

[0002] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application Serial No. 63 / 355,296 filed on June 24, 2022, U.S. Provisional Application Serial No. 63 / 370,484 filed on August 4, 2022, U.S. Provisional Application Serial No. 63 / 375,041 filed on September 8, 2022, and U.S. Provisional Application Serial No. 63 / 379,626 filed on October 14, 2022; the entire contents of each of these U.S. Provisional Applications are incorporated herein by reference.

[0003] Reference Electronic Sequence Listing

[0004] The contents of the electronic Sequence Listing (file name: B155370018WO00-SEQ-JRV.xml; size: 21,392 bytes; and creation date: June 19, 2023) are incorporated herein by reference in their entirety. Background Art

[0005] The complement system is a well-known effector mechanism of the immune response that provides not only protection against pathogens and other harmful substances, but also recovery from injury. The classical complement pathway is triggered by the activation of the first component of complement, called the C1 complex, which includes the C1q, C1r, and C1s proteins. After C1 binds to the immune complex, the C1s component, a diisopropylfluorophosphate (DFP)-sensitive serine protease, cleaves complement components C4 and C2 to initiate activation of the classical complement pathway. The classical complement pathway appears to play a role in many diseases and disorders, and safe and effective therapeutic agents targeting this pathway are needed. Summary of the invention

[0006] The present disclosure provides methods for treating classical complement-mediated disorders such as cold agglutinin disease (CAD) and chronic inflammatory demyelinating polyneuropathy (CIDP) using humanized antibodies that specifically bind to complement C1s protein (i.e., humanized anti-complement C1s antibodies, also referred to herein as "humanized anti-C1s antibodies," "humanized C1s antibodies," or "subject antibodies").

[0007] For classical complement-mediated disorders, such as CAD and CIDP, treatment options are limited. The subject antibody (a humanized monoclonal antibody to activated C1s) can provide clinical benefits to patients with classical complement system diseases by specifically targeting the activated classical pathway. The study described herein evaluates the safety, tolerability, and pharmacokinetic and pharmacodynamic profiles of the subject antibody in healthy participants and patients with CAD and CIDP. It is demonstrated herein that the subject antibody is generally well tolerated. The subject antibody shows acceptable bioavailability after SC administration, as well as a long elimination half-life with linear kinetics. The subject antibody further inhibits the activity of the classical complement pathway (CP) and 50% hemolytic complement activity (CH50) in a dose-dependent manner to levels consistent with a complement deficiency state. Preliminary results in CIDP patients further indicate that treatment with the subject antibody can improve the functional disability of CIDP patients who are refractory to other CIDP treatments and CIDP patients who are being treated with standard of care treatments, as measured by INCAT scores. The results support further evaluation of the subject antibodies in diseases characterized by dysfunctional or uncontrolled activation of the classical complement pathway, such as CAD and CIDP.

[0008] In one aspect, the present disclosure provides a method for treating cold agglutinin disease in a subject in need thereof, the method comprising administering to the subject about 3.5 g of a humanized antibody that specifically binds complement component C1s, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0009] In some embodiments, the antibody is administered about every 10, 12, 14, or 16 weeks. In some embodiments, the antibody is administered about every 12 weeks.

[0010] In some embodiments, about 3.5 g of the antibody is administered about 4 weeks after the first dose.

[0011] In some embodiments, the antibody is administered intravenously or subcutaneously. In some embodiments, the antibody is administered intravenously.

[0012] In some embodiments, about 3.5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29. In some embodiments, 3.5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional 3.5 g of the antibody is administered intravenously on day 29.

[0013] In some embodiments, about 3.5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29. In some embodiments, 3.5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and an additional 3.5 g of the antibody is administered intravenously on day 29.

[0014] In some embodiments, the administration of the antibody increases the level of hemoglobin in the subject. In some embodiments, the administration of the antibody reduces the level of bilirubin in the subject. In some embodiments, the administration of the antibody inhibits the activity of the classical complement pathway by about 90% or more. In some embodiments, the activity of the classical complement pathway is determined by measuring the amount of the terminal complement complex C5b-9. In some embodiments, the administration of the antibody reduces the level of 50% hemolytic complement activity (CH50) to less than about 10 IU / mL. In some embodiments, after the administration of the antibody, the subject has an antibody plasma concentration of at least about 100 μg / mL.

[0015] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 300 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0016] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 600 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0017] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 1200 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0018] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 2400 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0019] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 3600 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0020] In one aspect, the present disclosure provides a method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 7200 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, an LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0021] In some embodiments, one or more maintenance doses are administered approximately every week, every 2 weeks, every 4 weeks, or every 12 weeks.

[0022] In some embodiments, the antibody is administered intravenously or subcutaneously.

[0023] In some embodiments, a loading dose is administered intravenously. In some embodiments, one or more maintenance doses are administered subcutaneously.

[0024] In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose administered subcutaneously approximately weekly starting on day 8.

[0025] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 300 mg of the antibody administered subcutaneously about weekly starting on day 8. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 300 mg of the antibody administered subcutaneously about weekly starting on day 8.

[0026] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 600 mg of the antibody administered subcutaneously about weekly starting on day 8. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 600 mg of the antibody administered subcutaneously about weekly starting on day 8.

[0027] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 600 mg of the antibody administered subcutaneously about every 2 weeks starting on day 8. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 600 mg of the antibody administered subcutaneously every 2 weeks starting on day 8.

[0028] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 1200 mg of the antibody administered subcutaneously about every 2 weeks starting on day 8. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 1200 mg of the antibody administered subcutaneously every 2 weeks starting on day 8.

[0029] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 1200 mg of the antibody administered subcutaneously about every 4 weeks starting on day 29. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 1200 mg of the antibody administered subcutaneously every 4 weeks starting on day 29.

[0030] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 2400 mg of the antibody administered subcutaneously about every 4 weeks starting on day 29. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 2400 mg of the antibody administered subcutaneously every 4 weeks starting on day 29.

[0031] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 3600 mg of the antibody administered subcutaneously approximately every 12 weeks starting on day 29. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 3600 mg of the antibody administered subcutaneously every 12 weeks starting on day 29.

[0032] In some embodiments, a loading dose of about 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of about 7200 mg of the antibody administered subcutaneously approximately every 12 weeks starting on day 29. In some embodiments, a loading dose of 50 mg / kg of subject body weight of the antibody is administered intravenously on day 1, followed by a maintenance dose of 7200 mg of the antibody administered subcutaneously every 12 weeks starting on day 29.

[0033] In some embodiments, a one-time 50 mg / kg intravenous loading dose is administered on day 1, and a subcutaneous (SC) dose is administered with a prefilled syringe or autoinjector or a large volume drug delivery system. In some embodiments, the SC dose administered is 300 mg qw, 600 mg qw, 600 mg q2w, 1200 mg q2w, 1200 mg q4w, 2400 mg q4w, 3600 mgq12w, or 7200 mg q12w. In some embodiments, a SC dose is administered with a prefilled syringe or autoinjector or a large volume drug delivery system until the end of treatment.

[0034] In some embodiments, the subject is refractory to another CIDP treatment. In some embodiments, the subject has received another CIDP treatment within about one week of the loading dose, or simultaneously with another CIDP treatment. In some embodiments, the subject has not received another CIDP treatment within about 6 months prior to the loading dose. In some embodiments, the another CIDP treatment is intravenous immunoglobulin (IVIg), subcutaneous immunoglobulin (SCIg), or corticosteroids.

[0035] In some embodiments, administration of the antibody results in a decrease in adjusted Inflammatory Neuropathy Cause and Treatment (INCAT) disability score of one point or more relative to the INCAT score prior to treatment with the antibody.

[0036] In some embodiments, following administration of the antibody, the subject has a plasma concentration of the antibody of at least about 1200 μg / mL.

[0037] In some embodiments, the antibody is contained in a syringe or vial.

[0038] In some embodiments, the antibody is administered using a syringe, pre-filled syringe, or a large volume drug delivery system.

[0039] In some embodiments, the antibody is administered using an automatic injector.

[0040] In some embodiments, the antibody is administered using a large volume device for subcutaneous injection.

[0041] In some embodiments, the antibody comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:8.

[0042] In some embodiments, the antibody is a Fab fragment, a F(ab')2 fragment, a scFv, or a Fv.

[0043] In some embodiments, the antibody comprises a heavy chain constant region of isotype IgG4.

[0044] In some embodiments, the IgG4 constant region comprises proline, glutamic acid, leucine, and serine substitutions at amino acid residues 108, 115, 308, and 314, respectively, relative to the IgG4 constant region sequence of SEQ ID NO: 11.

[0045] In some embodiments, the IgG4 constant region comprises the sequence of SEQ ID NO:13.

[0046] In some embodiments, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:9 and a heavy chain comprising the amino acid sequence of SEQ ID NO:10.

[0047] Each of International Publication Nos. WO 2014 / 066744, filed on October 25, 2013, entitled “Anti-Complement C1s Antibodies and Uses Thereof,” WO 2016 / 164358, filed on April 5, 2016, entitled “Humanized Anti-C1s Antibodies and Methods of Use Thereof,” and WO 2018 / 071676, filed on October 12, 2017, entitled “Anti-C1s Antibodies and Methods of Use Thereof,” are incorporated herein by reference in their entirety. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 Schematic diagram showing the disposition of participants. IV, intravenous; SC, subcutaneous; PD, pharmacodynamic; PK, pharmacokinetic; *Japanese.

[0049] Figures 2A-2D Pharmacokinetic (mean ± SD) plasma concentration-time profiles are shown: (a) linear scale single dose IV cohort (mean ± SD); (b) linear scale single dose SC cohort (mean ± SD); (c) multiple doses 300 mg or 600 mg SC after the last weekly dose; (d) daily multiple doses 600 mg SC for three doses. IV, intravenous; h, hour; SC, subcutaneous; SD, standard deviation.

[0050] Figures 3A-3F Pharmacodynamic effects on the classical complement pathway and CH50 are shown (Part 1 and Part 2). (a) CP inhibition percentage after a single IV dose - time curve. (b) CP inhibition percentage after a single SC dose - time curve. (c) CP inhibition percentage after repeated SC doses - time curve. (d) CH50 - time curve after a single IV dose. (e) CH50 - time curve after a single SC dose. (f) CH50 - time curve after repeated SC doses. CP, classical complement pathway; CH50, 50% hemolytic complement activity; IV, intravenous; SC, subcutaneous; SEM, standard error of the mean.

[0051] Figures 4A-4E Pharmacokinetic / pharmacodynamic correlations are shown: CP percentage, CH50, AP percentage, and C4 profile. (a) Visual predictive check of the final healthy subject population PK model. Note: Legend: Blue dots: observations; solid and red dashed lines: median and limit values ​​(5th and 95th percentiles) of observed concentrations in each time zone; pink and light blue areas: confidence intervals for median and percentile predicted concentrations in each time zone. The X-axis is the time after administration of the first dose. (b) CH50 vs. SAR445088 (single dose) concentration for the IV cohort. (c) CH50 vs. SAR445088 (single dose) concentration curves for the SC dose. (d) % CP inhibition vs. SAR445088 (single dose) concentration for the IV cohort. (e) % CP inhibition vs. SAR445088 (single dose) concentration for the SC dose. AP, alternative pathway; CH50, 50% hemolytic complement activity; CP, classical complement pathway; IV, intravenous; SC, subcutaneous.

[0052] Figure 5Summary graph of total bilirubin (umol / L) versus baseline over time - Biomarker Activity (BA) population is shown. Baseline is defined as D1-1H assessment. D = day; H = hour.

[0053] Figure 6 Summary graph showing changes in hemoglobin (g / L) relative to baseline over time - BA population. Baseline is defined as D1-1H assessment. D = day; H = hour.

[0054] Figure 7 Shown are the observed SAR445088 concentration-time profiles in CAD patients and the 90% prediction intervals of the popPK model based on healthy subject data.

[0055] Figure 8 Shown is a PK / PD simulation of the SAR445088 regimen used in the Phase 2 CAD study: 30 mg / kg IV.

[0056] Fig. 9 PK / PD simulation of the SAR445088 regimen for the Phase 2 CIDP study is shown: 50 mg / kg IV on Day 1 + 600 mg SC qw from Day 8. Red line: predicted SAR445088 concentration, red shaded 90% prediction interval. Blue line: predicted CH50 level; blue shaded 90% prediction interval.

[0057] Fig.10 PopPK simulation of a flat dose regimen is shown: 3.5 g IV q12w starting on day 1 + 3.5 g IV on day 29.

[0058] Fig.11 Shown are box plots of SAR445088 constant dose versus weight-based dose simulated steady-state AUC.

[0059] Fig.12 Shown is a constant dose of SAR445088 versus a weight-based dose to simulate steady-state C trough Box plot.

[0060] Fig.13 A schematic diagram of the Phase 2 CIDP study is shown.

[0061] Fig.14 An overview of all efficacy endpoints in all trial patients - the primary endpoint INCAT score and the secondary efficacy endpoints I-RODS total score, grip strength and MRC total score (MRCSS) are shown. The overall data are shown as mean ± SD relative to the change of baseline. The number of patients contributing to each time point from each subgroup is in the table below the chart.

[0062] Fig.15 INCAT score, SOC refractory patients, longitudinal changes of group data relative to baseline are shown. Data include all available patients / time points at the time of cutoff. Error bars indicate standard deviation (SD).

[0063] Fig.16 Longitudinal changes from baseline in INCAT scores, SOC-treated patients, and group data are shown.

[0064] Fig.17 Longitudinal changes from baseline in INCAT scores, patients not receiving SOC, group data are shown.

[0065] Fig.18 Complement inhibition (CH50 biomarker) in patients refractory to SOC (N=16), treated with SOC (N=22), and not receiving SOC (N=6) is shown. The dashed line represents the expected level of complement deficiency based on sultilimab (<10 IU / ml cutoff). Data include all available patients / time points at the time of cutoff. Error bars represent SD.

[0066] Fig.19 Shown are changes from baseline in key clinical outcome measures evaluating functional disability (INCAT disability score, I-RODS) and impairment (grip strength, MRC-SS), SOC refractory group (N=12), interim analysis at 50% total sample size. The dotted line represents the minimal clinically important difference (MCID) cutoff value established in the field (2021EAN / PNS guidelines).

[0067] Fig. 20 Shown are changes from baseline in key clinical outcome measures evaluating functional disability (INCAT disability score, I-RODS) and impairment (grip strength, MRC-SS), SOC treatment group (N=25), interim analysis at 50% total sample size. The dotted line represents the minimal clinically important difference (MCID) cutoff value established in the field (2021EAN / PNS guidelines).

[0068] Figures 21A-21B Shown are changes in serum NfL levels from baseline. Fig.21A .Overall data for the three study groups. Fig.21B . Detailed and extended analyses in response-based SOC treatment groups. DETAILED DESCRIPTION

[0069] Humanized anti-C1s antibody

[0070] The methods of the present disclosure include administering a humanized anti-C1s antibody. The humanized anti-C1s antibody of the present disclosure binds to and inhibits the activated form of C1s within the classical pathway (CP). The complement system is a component of the innate immune system that mediates humoral immunity. The mechanism of the antibody is specific for the CP, thereby leaving the lectin and alternative pathways functionally intact.

[0071] In some embodiments, the humanized anti-C1s antibodies of the present disclosure inhibit C1s-mediated cleavage of complement component C4, for example, by inhibiting the enzymatic activity of the serine-protease domain of C1s. In some embodiments, the humanized anti-C1s antibodies of the present disclosure inhibit C1s-mediated cleavage of complement component C2. In some embodiments, the humanized anti-C1s antibodies of the present disclosure inhibit C1s-mediated cleavage of C4 and C2.

[0072] In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to a complement C1s protein having the amino acid sequence depicted in SEQ ID NO: 15. The amino acid sequence SEQ ID NO: 15 provided below represents the Homo sapiens complement C1s protein:

[0073] EPTMYGEILSPNYPQAYPSEVEKSWDIEVPEGYGIHLYFTHLDIELSENCA

[0074] YDSVQIISGDTEEGRLCGQRSSNNPHSPIVEEFQVPYNKLQVIFKSDFSNEE

[0075] RFTGFAAYYVATDINECTDFVDVPCSHFCNNFIGGYFCSCPPEYFLHDDM

[0076] KNCGVNCSGDVFTALIGEIASPNYPKPYPENSRCEYQIRLEKGFQVVVTL

[0077] RREDFDVEAADSAGNCLDSLVFVAGDRQFGPYCGHGFPGPLNIETKSNA

[0078] LDIIFQTDLTGQKKGWKLRYHGDPMPCPKEDTPNSVWEPAKAKYVFRD

[0079] VVQITCLDGFEVVEGRVGATSFYSTCQSNGKWSNSKLKCQPVDCGIPESI

[0080] ENGKVEDPESTLFGSVIRYTCEEPYYYMENGGGGEYHCAGNGSWVNEV

[0081] LGPELPKCVPVCGVPREPFEEKQRIIGGSDADIKNFPWQVFFDNPWAGGA

[0082] LINEYWVLTAAHVVEGNREPTMYVGSTSVQTSRLAKSKMLTPEHVFIHP

[0083] GWKLLEVPEGRTNFDNDIALVRLKDPVKMGPTVSPICLPGTSSDYNLMD

[0084] GDLGLISGWGRTEKRDRAVRLKAARLPPVAPLRKCKEVKVEKPTADAEA

[0085] YVFTPNMICAGGEKGMDSCKGDSGGAFAVQDPNDKTKFYAAGLVSWGPQCGTYGLYTRVKNYVDWIMKTMQENSTPRED (SEQ ID NO: 15).

[0086] In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a dissociation constant (KD) of no more than 2.5 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 2 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 1 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 0.9 nM, no more than 0.8 nM, no more than 0.7 nM, no more than 0.6 nM, no more than 0.5 nM, no more than 0.4 nM, no more than 0.3 nM, no more than 0.2 nM or no more than 0.1 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 0.3 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 0.2 nM. In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 0.1 nM. Methods for measuring the binding of antibodies to human complement C1s protein can be determined by those skilled in the art.

[0087] In some embodiments, the humanized anti-C1s antibodies of the present disclosure bind to human complement C1s protein with a KD of no more than 90 pM, no more than 80 pM, no more than 70 pM, no more than 60 pM, no more than 50 pM, no more than 40 pM, no more than 30 pM, no more than 20 pM, no more than 10 pM, no more than 9 pM, no more than 8 pM, no more than 7 pM, no more than 6 pM, no more than 5 pM, no more than 4 pM, no more than 3 pM, no more than 2 pM, or no more than 1 pM.

[0088] In some embodiments, the humanized anti-C1s antibodies of the present disclosure preferentially bind to an active (or activated) human complement C1s protein and do not substantially bind to an inactive form of the human complement C1s protein.

[0089] In some embodiments, the humanized anti-C1s antibodies disclosed herein are -8 M or lower, 5×10 -9 M or lower, or 10 -9 M or lower half maximal inhibitory concentration (IC 50 ) inhibits the classical complement pathway.

[0090] "Antibodies" encompass antibodies or immunoglobulins of any isotype, including but not limited to humanized antibodies and chimeric antibodies. Antibodies can be single-chain antibodies (scAbs) or single-domain antibodies (dAbs) (e.g., single-domain heavy chain antibodies or single-domain light chain antibodies; see Holt et al. (2003) Trends Biotechnol. [Biotech Trends] 21:484). The term "antibody" also encompasses fragments of antibodies that retain specific binding to an antigen (antibody fragments). "Antibodies" further include: single-chain variable fragments (scFv), which are antibody heavy chains (V H ) and light chain (V L ) fusion protein of the variable region of the scFv fragment, connected to a short linker peptide; and a diabody, which is a non-covalent dimer of the scFv fragment, including the V H and V L (Zapata et al., Protein Eng. 8(10):1057-1062 (1995)) Other fusion proteins comprising the antigen-binding portion of an antibody and a non-antibody protein are also encompassed by the term "antibody."

[0091] "Antibody fragments" include a portion of an intact antibody, such as the antigen binding region or variable region of an intact antibody. Examples of antibody fragments include antigen binding fragments (Fab), Fab', F(ab')2, variable domain Fv fragments (Fv), Fd fragments, and antigen binding fragments of chimeric antigen receptors.

[0092] Papain digestion of antibodies produces: two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, a name reflecting the ability to crystallize readily. Pepsin treatment yields the F(ab')2 fragment, which has two antigen-binding sites and is still capable of cross-linking antigen.

[0093] "Fv" is the smallest antibody fragment that contains a complete antigen recognition and antigen binding site. This region includes a dimer of one heavy chain variable domain and one light chain variable domain in tight, non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define V H -V L The antigen binding site on the surface of the dimer. The six CDRs together give the antibody antigen binding specificity. However, even a single variable domain (or half of an Fv that includes only three CDRs specific for an antigen) has the ability to recognize and bind to an antigen, although with a lower affinity than the complete binding site.

[0094] The "Fab" fragment contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain containing at least one cysteine ​​from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine ​​residues of the constant domains bear free thiol groups. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0095] The "scFv" antibody fragment contains the V H and V L , wherein these regions are present in a single polypeptide chain. In some embodiments, the Fv polypeptide further comprises a H Area and V L The polypeptide linker between the two regions enables the scFv to form the desired structure for antigen binding. For a review of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0096] "Diabodies" are small antibody fragments with two antigen-binding sites. These fragments contain two antigen-binding sites on the same polypeptide chain (V H -VL ) L Connect V H By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites. Diabodies are more fully described in, for example, Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993).

[0097] Antibodies may be monovalent or divalent.Antibodies may be Ig monomers, which are "Y-shaped" molecules composed of four polypeptide chains: two heavy chains and two light chains linked by disulfide bonds.

[0098] The antibody can be detectably labeled, for example, with a radioisotope, an enzyme that generates a detectable product, and / or a fluorescent protein. The antibody can be further conjugated to other moieties, such as members of a specific binding pair, e.g., the biotin member of a biotin-avidin specific binding pair. The antibody can also be bound to a solid support, including but not limited to polystyrene plates and / or beads.

[0099] "Isolated" antibodies are antibodies that have been identified and separated and / or recovered from components of their natural environment (i.e., not naturally occurring). The contaminating components of their natural environment are substances that interfere with the use of the antibody (e.g., diagnostic or therapeutic use), and may include enzymes, hormones, and other proteins or non-protein solutes. In some embodiments, the antibody is purified (1) to an antibody greater than 90% by weight, greater than 95% by weight, greater than 98% by weight, or greater than 99% by weight as determined by, for example, the Lowry method, (2) to a degree sufficient to obtain at least 15 residues or internal amino acid sequences of the N-terminus using a spinning cup sequencer, or (3) to homogeneity obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing or non-reducing conditions using Coomassie blue or silver staining. The isolated antibody encompasses antibodies in situ within a recombinant cell, because at least one component of the antibody's natural environment will not be present. In some embodiments, the isolated antibody is prepared by at least one purification step.

[0100] "Monoclonal antibody" is an antibody produced by a group of identical cells, all of which are produced from a single cell by repeated cell replication. That is, the cell clone produces only a single antibody species. Although monoclonal antibodies can be produced using hybridoma production technology, other production methods known to those skilled in the art (e.g., antibodies derived from antibody phage display libraries) can also be used.

[0101] "Complementarity determining regions (CDRs)" are non-contiguous antigen combining sites present in the variable regions of both heavy and light chain polypeptides. CDRs have been described by Lefranc et al. (2003) Developmental and Comparative Immunology 27:55; Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., USDept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987); and MacCallum et al., J. Mol. Biol. 262:732-745 (1996), wherein these definitions encompass overlapping or subsets of amino acid residues when compared to each other. However, the use of any of the Kabat, Lefranc, Chothia or MacCallum definitions (also referred to as numbering systems) to refer to the CDRs of an antibody or grafted antibody or variants thereof is intended to be within the scope of the term as defined and used herein.

[0102] The terms "LC CDR1", "LC CDR2" and "LC CDR3" refer to the first CDR, the second CDR and the third CDR in the light chain variable region, respectively. As used herein, the terms "HC CDR1", "HC CDR2" and "HC CDR3" refer to the first CDR, the second CDR and the third CDR in the heavy chain variable region, respectively. As used herein, the terms "CDR1", "CDR2" and "CDR3" refer to the first CDR, the second CDR and the third CDR of the variable region of either chain, respectively.

[0103] When used to refer to an antibody variable region, "framework" includes all amino acid residues outside the CDR region within the antibody variable region. The variable region framework is typically a discontinuous amino acid sequence that includes only those amino acids outside the CDR. A "framework region" includes each domain of the framework separated by a CDR.

[0104] "Humanized antibody" is an antibody comprising parts of antibodies from different sources, wherein at least one part comprises an amino acid sequence of human origin. For example, a humanized antibody may comprise parts of antibodies from non-human sources (such as mice) with the necessary specificity and parts of antibody sequences (e.g., chimeric immunoglobulins) from human origin, which are chemically linked together by conventional techniques (e.g., synthesis) or prepared as continuous polypeptides using genetic engineering techniques (e.g., DNA encoding the protein portion of a chimeric antibody can be expressed to produce a continuous polypeptide chain). Another example of a humanized antibody is an antibody containing at least one chain, which comprises a CDR derived from an antibody from a non-human source and a framework region of a light chain and / or a heavy chain derived from a human source (e.g., a CDR-grafted antibody with or without a framework change). The term humanized immunoglobulin also encompasses chimeric or CDR-grafted single-chain antibodies. See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; Cabilly et al., European Patent No. 0,125,023 B1; Boss et al., U.S. Pat. No. 4,816,397; Boss et al., European Patent No. 0,120,694 B1; Neuberger, MS et al., WO 86 / 01533; Neuberger, MS et al., European Patent No. 0,194,276 B1; Winter, U.S. Pat. No. 5,225,539; Winter, European Patent No. 0,239,400 B1; Padlan, EA et al., European Patent Application No. 0,519,596 A1. For single-chain antibodies, see also Ladner et al., U.S. Pat. No. 4,946,778; Huston, U.S. Pat. No. 5,476,786; and Bird, RE et al., Science, 242:423-426 (1988).

[0105] In some embodiments, humanized antibodies are produced using synthetic and / or recombinant nucleic acids to prepare genes (e.g., cDNA) encoding the desired humanized chains. For example, nucleic acid (e.g., DNA) sequences encoding humanized variable regions can be constructed using PCR mutagenesis methods to change the DNA sequence encoding a human or humanized chain, such as a DNA template from a previously humanized variable region (see, e.g., Kamman, M. et al., Nucl. Acids Res. [Nucleic Acids Research], 17: 5404 (1989); Sato, K. et al., Cancer Research [Cancer Research], 53: 851-856 (1993); Daugherty, BL et al., Nucleic Acids Res. [Nucleic Acids Research], 19 (9): 2471-2476 (1991); and Lewis, AP and JS Crowe, Gene [Gene], 101: 297-302 (1991)). Variants can also be easily generated using these or other suitable methods. For example, the cloned variable regions can be subjected to mutagenesis and sequences encoding variants with the desired specificity can be selected (e.g., from phage libraries; see, e.g., Krebber et al., U.S. Pat. No. 5,514,548; Hoogenboom et al., WO 93 / 06213, published April 1, 1993).

[0106] In some embodiments, the humanized anti-C1s antibodies described herein are full-length IgG, Ig monomers, Fab fragments, F(ab')2 fragments, Fd fragments, scFv, scAb or Fv. In some embodiments, the humanized anti-C1s antibodies described herein are full-length IgG. In some embodiments, the heavy chain of any humanized anti-C1s antibody as described herein comprises a heavy chain constant region (CH) or a portion thereof (e.g., CH1, CH2, CH3 or a combination thereof). The heavy chain constant region can be of any suitable origin, such as human, mouse, rat or rabbit. In some embodiments, the heavy chain constant region is from human IgG (gamma heavy chain), such as IgG1, IgG2 or IgG4.

[0107] In some embodiments, mutations can be introduced into the heavy chain constant region of any of the humanized anti-C1s antibodies described herein. In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the heavy chain constant region (e.g., CH2 domain (residues 231-340 of human IgG1) and / or CH3 domain (residues 341-447 of human IgG1) and / or hinge region, numbered according to the Kabat numbering system (e.g., EU index in Kabat)) to increase or decrease the affinity of the antibody for an Fc receptor on the surface of an effector cell (e.g., an activated Fc receptor). Mutations in the Fc region of an antibody that decrease or increase the affinity of the antibody for an Fc receptor and techniques for introducing such mutations into an Fc receptor or fragment thereof are known to those of skill in the art. Examples of mutations that can be made in the Fc receptor of an antibody to alter the affinity of the antibody for the Fc receptor are described in, e.g., Smith P et al., (2012) PNAS [Proceedings of the National Academy of Sciences of the United States of America] 109:6181-6186; U.S. Pat. No. 6,737,056 and International Publication Nos. WO 02 / 060919, WO 98 / 23289 and WO 97 / 34631, which are incorporated herein by reference.

[0108] In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the heavy chain constant region (CH1 domain) such that the number of cysteine ​​residues in the hinge region is altered (e.g., increased or decreased), as described, e.g., in U.S. Pat. No. 5,677,425. The number of cysteine ​​residues in the hinge region of the CH1 domain may be altered, e.g., to facilitate assembly of the light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody or to facilitate linker conjugation.

[0109] In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into IgG constant domains or their FcRn binding fragments to change (e.g., shorten or increase) the in vivo half-life of the antibody. In some embodiments, one or more mutations are introduced into Fc or hinge-Fc domain fragments. For examples of mutations that will change (e.g., shorten or increase) the in vivo half-life of an antibody, see, e.g., International Publication Nos. WO 02 / 060919, WO 98 / 23289 and WO 97 / 34631 and U.S. Patent Nos. 5,869,046, 6,121,022, 6,277,375 and 6,165,745.

[0110] In some embodiments, the constant region antibodies described herein are IgG1 constant regions and comprise a methionine (M)-tyrosine (Y) substitution at position 252, a serine (S)-threonine (T) substitution at position 254, and a threonine (T)-glutamic acid (E) substitution at position 256, numbered according to the EU index in Kabat. See U.S. Patent No. 7,658,921, which is incorporated herein by reference. This type of mutant IgG, referred to as a "YTE mutant," has been shown to exhibit a four-fold increase in half-life compared to the wild-type version of the same antibody (see Dall'Acqua WF et al., (2006) J Biol Chem [Journal of Biological Chemistry] 281: 23514-24). In some embodiments, the antibody comprises an IgG constant domain comprising one, two, three or more amino acid substitutions of amino acid residues at positions 251-257, 285-290, 308-314, 385-389 and 428-436 numbered according to the EU index in Kabat. Additional mutations that can be introduced into the heavy chain constant region to increase antibody half-life are known in the art, such as the M428L / N434S (EU numbering; M459L / N466S Kabat numbering) mutations as described in Zalevsky et al., Nat Biotechnol. 2010 Feb;28(2):157–159.

[0111] In some embodiments, one, two or more amino acid substitutions are introduced into the IgG constant domain Fc region to change the effector function of the antibody. The effector ligand with changed affinity can be, for example, an Fc receptor or the C1 component of complement. This method is further described in detail in U.S. Patent Nos. 5,624,821 and 5,648,260. In some embodiments, the deletion or inactivation of the constant region domain (by point mutation or other means) can reduce the Fc receptor binding of circulating antibodies, thereby increasing tumor localization. For descriptions of mutations that delete or inactivate the constant domain and thereby increase tumor localization, see, for example, U.S. Patent Nos. 5,585,097 and 8,591,886. In some embodiments, at least one amino acid substitution can be introduced into the Fc region of the antibody described herein to remove potential glycosylation sites on the Fc region, thereby reducing Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem [Journal of Biological Chemistry] 276: 6591-604).

[0112] In some embodiments, at least one amino acid in the constant region can be replaced by a different amino acid residue so that the antibody has altered C1q binding and / or reduced or eliminated complement dependent cytotoxicity (CDC). This method is further described in detail in U.S. Patent No. 6,194,551 (Idusogie et al.). In some embodiments, at least one amino acid residue in the N-terminal region of the CH2 domain of the antibodies described herein is altered, thereby changing the ability of the antibody to fix complement. This method is further described in International Publication No. WO 94 / 29351. In some embodiments, the Fc region of the antibodies described herein is modified to increase the ability of the antibody to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or increase the affinity of the antibody for Fcγ receptors. This method is further described in International Publication No. WO 00 / 42072.

[0113] In some embodiments, to avoid potential complications due to Fab-arm exchange known to occur with native IgG4 mAbs, the antibodies provided herein may include a stabilizing "Adair" mutation (Angal S. et al., "A single aminoacid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody", Mol Immunol 30, 105-108; 1993), in which serine 228 (EU numbering; residue 241 Kabat numbering) is converted to proline, thereby generating an IgG1-like hinge sequence. In some embodiments, to reduce residual antibody-dependent cellular toxicity, an L235E (EU numbering, corresponding to L248E in Kabat numbering) mutation is introduced into the heavy chain constant region, e.g., as described in Benhnia et al., J. Virology, December 2009, pp. 12355-12367.

[0114] In some embodiments, the humanized anti-C1s antibody comprises a light chain complementary determining region 1 (LC CDR1) comprising the amino acid sequence KASQSVDYDGDSYMN (SEQ ID NO: 1). In some embodiments, the humanized anti-C1s antibody comprises a light chain complementary determining region 2 (LC CDR2) comprising the amino acid sequence DASNLES (SEQ ID NO: 2). In some embodiments, the humanized anti-C1s antibody comprises a light chain complementary determining region 3 (LC CDR3) comprising the amino acid sequence QQSNEDPWT (SEQ ID NO: 3). In some embodiments, the humanized anti-C1s antibody comprises a LC CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3.

[0115] In some embodiments, the humanized anti-C1s antibody comprises a heavy chain complementary determining region 1 (HC CDR1) comprising the amino acid sequence DDYIH (SEQ ID NO:4). In some embodiments, the humanized anti-C1s antibody comprises a heavy chain complementary determining region 2 (HC CDR2) comprising the amino acid sequence RIDPADGHTKYAPKFQV (SEQ ID NO:5). In some embodiments, the humanized anti-C1s antibody comprises a heavy chain complementary determining region 3 (HC CDR3) comprising the amino acid sequence YGYGREVFDY (SEQ ID NO:6). In some embodiments, the humanized anti-C1s antibody comprises a HC CDR1 comprising the amino acid sequence of SEQ ID NO:4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a HC CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0116] In some embodiments, the humanized anti-C1s antibody comprises a LC CDR1 comprising the amino acid sequence of SEQ ID NO:1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO:2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO:3, a HC CDR1 comprising the amino acid sequence of SEQ ID NO:4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a HC CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0117] In some embodiments, the humanized anti-C1s antibody comprises a light chain variable region (VL) comprising the amino acid sequence DIVLTQSPDSLAVSLGERATISCKASQSVDYDGD SYMNWYQQKPGQPPKILIYDASNLESGIPARFSGSGSGTDFTLTISSLEPED FAIYYCQQSNEDPWTFGGGTKVEIK (SEQ ID NO: 7). In some embodiments, the humanized anti-C1s antibody comprises a LC CDR1, LC CDR2, and LC CDR3 of a VL comprising the amino acid sequence of SEQ ID NO: 7.

[0118] In some embodiments, the humanized anti-C1s antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence QVQLVQSGAEVKKPGASVKLSCTASGFNIKDDY IHWVKQAPGQGLEWIGRIDPADGHTKYAPKFQVKVTITADTSTSTAYLELSSLRSEDTAVYYCARYGYGREVFDYWGQGTTVTVSS (SEQ ID NO: 8). HC CDR1, HC CDR2, and HC CDR3 of the VH comprising the amino acid sequence of SEQ ID NO: 8.

[0119] In some embodiments, the humanized anti-C1s antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:7 and a VH comprising the amino acid sequence of SEQ ID NO:8.

[0120] In some embodiments, the humanized anti-C1s antibody comprises LC CDR1, LC CDR2, and LC CDR3 of VL comprising the amino acid sequence of SEQ ID NO:7, and HC CDR1, HC CDR2, and HC CDR3 of VH comprising the amino acid sequence of SEQ ID NO:8.

[0121] In some embodiments, the humanized anti-C1s antibody comprises a light chain (LC) comprising the amino acid sequence DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQKPGQPPKILIYDASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAIYYCQQSNEDPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 9).

[0122] In some embodiments, the humanized anti-C1s antibody comprises a heavy chain (HC) comprising the amino acid sequence QVQLVQSGAEVKKPGASVKLSCTASGFNIKDDYIHWVKQAP GQGLEWIGRIDPADGHTKYAPKFQVKVTITADTSTSTAYLELSSLRSEDTAVYYCARYGYGREVFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQV YTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHSHYTQKSLSLSLGK(SEQ ID NO:10).

[0123] In some embodiments, the humanized anti-C1s antibody comprises a LC comprising the amino acid sequence of SEQ ID NO:9 and a HC comprising the amino acid sequence of SEQ ID NO:10.

[0124] In some embodiments, the humanized anti-C1s antibody comprises a LC CDR1 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the LC CDR1 amino acid sequence of SEQ ID NO: 1. In some embodiments, the humanized anti-C1s antibody comprises a LC CDR2 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the LC CDR2 amino acid sequence of SEQ ID NO: 2. In some embodiments, the humanized anti-C1s antibody comprises a LC CDR3 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the LC CDR3 amino acid sequence of SEQ ID NO: 3.

[0125] In some embodiments, the humanized anti-C1s antibody comprises a HC CDR1 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the HC CDR1 amino acid sequence of SEQ ID NO: 4. In some embodiments, the humanized anti-C1s antibody comprises a HC CDR2 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the HC CDR2 amino acid sequence of SEQ ID NO: 5. In some embodiments, the humanized anti-C1s antibody comprises a HC CDR3 comprising an amino acid sequence containing no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) relative to the HC CDR3 amino acid sequence of SEQ ID NO: 6.

[0126] In some embodiments, the humanized anti-C1s antibody comprises a VL comprising an amino acid sequence that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) relative to the VL amino acid sequence of SEQ ID NO:7.

[0127] In some embodiments, the humanized anti-C1s antibody comprises a VH comprising an amino acid sequence that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) relative to the VH amino acid sequence of SEQ ID NO:8.

[0128] In some embodiments, the humanized anti-C1s antibody comprises a VL comprising a LC CDR1 comprising the amino acid sequence of SEQ ID NO:1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO:2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO:3, and framework regions containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) relative to the VL sequence of SEQ ID NO:7.

[0129] In some embodiments, the humanized anti-C1s antibody comprises a VH comprising a HC CDR1 comprising the amino acid sequence of SEQ ID NO:4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO:5, a HC CDR3 comprising the amino acid sequence of SEQ ID NO:6, and framework regions containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) relative to the VH sequence of SEQ ID NO:8.

[0130] In some embodiments, the humanized anti-C1s antibody comprises: (a) a VL comprising a LC CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and comprising framework regions that contain no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) relative to the VL sequence of SEQ ID NO: 7; and (b) a VH comprising a HC CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6, and comprising framework regions that contain no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) relative to the VL sequence of SEQ ID NO: The VH sequence of NO:8 contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation).

[0131] In some embodiments, the humanized anti-C1s antibody comprises a VL comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to the VL amino acid sequence of SEQ ID NO:7.

[0132] In some embodiments, the humanized anti-C1s antibody comprises a VH comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to the VH amino acid sequence of SEQ ID NO:8.

[0133] In some embodiments, the humanized anti-C1s antibody comprises a VL comprising a LC CDR1 comprising the amino acid sequence of SEQ ID NO:1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO:2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO:3, and framework regions that are at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to the framework regions of the VL sequence of SEQ ID NO:7.

[0134] In some embodiments, the humanized anti-C1s antibody comprises a VH comprising a HC CDR1 comprising the amino acid sequence of SEQ ID NO:4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO:5, a HC CDR3 comprising the amino acid sequence of SEQ ID NO:6, and framework regions that are at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to the framework regions of the VH sequence of SEQ ID NO:8.

[0135] In some embodiments, the humanized anti-C1s antibody comprises: (a) a VL comprising a LC CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and framework regions that are at least 80% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) identical to the framework regions of the VL sequence of SEQ ID NO: 7; and (b) a VH comprising a HC CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HC CDR3 comprising the amino acid sequence of SEQ ID NO: 6, and framework regions that are identical to the framework regions of the VL sequence of SEQ ID NO: 7. The framework regions of the VH sequence of NO:8 have at least 80% (eg, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) identity.

[0136] In some embodiments, the heavy chain constant region in any of the humanized anti-C1s antibodies described herein is an IgG4 constant region, or a variant thereof. Examples of IgG4 constant regions and variants are provided in Table 1.

[0137] Table 1. Examples of heavy chain constant regions

[0138]

[0139] In some embodiments, the light chain of any of the humanized anti-C1s antibodies described herein may further comprise a light chain constant region (C L ). In some examples, C L is a kappa light chain. In other embodiments, C L is a lambda light chain. In some embodiments, C L is a kappa light chain, the sequence of which is provided below: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC (SEQ ID NO: 14)

[0140] Other antibody heavy and light chain constant regions are well known in the art, for example, those provided in the IMGT database (imgt.org) or vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0141] Treatment options for CAD

[0142] Cold agglutinin disease (CAD) is an autoimmune hemolytic anemia caused by IgM-induced activation of the classical complement pathway. The IgM autoantibodies in CAD are called cold agglutinins (CAgs) because of their inherent property of increasing binding when the temperature is reduced. In many patients, the CAg thermal amplitude is close to the core body temperature, which is the highest temperature at which CAg can be detected reacting with red blood cell antigens. Symptoms of CAD are usually triggered by exposure to cold ambient temperatures (i.e., temperatures at or below core body temperature), viral infection, or inflammation. All patients with CAD have active disease with varying levels of chronic, persistent hemolysis, resulting in anemia.

[0143] The present disclosure provides methods for treating CAD in a subject. In one aspect, these methods comprise administering an anti-C1s antibody to a subject at a fixed (or constant) dose of about 3 g to about 5 g. In some embodiments, these methods comprise administering an anti-C1s antibody to a subject at a fixed dose of about 3.5 g. In some embodiments, these methods comprise administering an anti-C1s antibody to a subject at a fixed dose of about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 4.5 g.

[0144] In some embodiments, the antibody is administered about every 10, 12, 14, or 16 weeks. In some embodiments, the anti-C1s antibody is administered about every 12 weeks.

[0145] In some embodiments, an additional anti-C1s antibody is administered about 4 weeks after the first dose. In some embodiments, about 3 g to about 5 g of anti-C1s antibody is administered about 4 weeks after the first dose. In some embodiments, about 3.5 g of anti-C1s antibody is administered about 4 weeks after the first dose. In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 4.5 g of anti-C1s antibody is administered about 4 weeks after the first dose.

[0146] In some embodiments, about 3 g to about 5 g of the antibody is administered on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3 g to about 5 g of the antibody is administered on or about day 29. In some embodiments, about 3 g to about 5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3 g to about 5 g of the antibody is administered intravenously on or about day 29.

[0147] In some embodiments, about 3 g to about 5 g of the antibody is administered on day 1 and about every 12 weeks thereafter, and about 3 g to about 5 g of the antibody is additionally administered on or about day 29. In some embodiments, about 3 g to about 5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and about 3 g to about 5 g of the antibody is additionally administered intravenously on or about day 29.

[0148] In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered on or about day 29. In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered intravenously on or about day 29.

[0149] In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered on day 1 and about every 12 weeks thereafter, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered on or about day 29. In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered intravenously on or about day 29.

[0150] In some embodiments, about 3.5 g of the antibody is administered on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3.5 g of the antibody is administered on or about day 29. In some embodiments, about 3.5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29.

[0151] In some embodiments, about 3.5 g of the antibody is administered on day 1 and about every 12 weeks thereafter, and an additional about 3.5 g of the antibody is administered on or about day 29. In some embodiments, about 3.5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29.

[0152] In some embodiments, about 3 g to about 5 g of anti-C1s antibody is administered to a subject about every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year. In some cases, about 3 g to about 5 g of anti-C1s antibody is administered to a subject about every twelve weeks for a period of more than 1 year. For example, in some embodiments, about 3 g to about 5 g of anti-C1s antibody is administered to a subject about every 12 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0153] In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of anti-C1s antibody is administered to a subject about every 12 weeks for a period of about 12 weeks to 1 year, such as about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year. In some cases, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of anti-C1s antibody is administered to a subject every twelve weeks for a period of more than 1 year. For example, in some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of anti-C1s antibody is administered to a subject about every 12 weeks for a period of 1 to 50 years, such as 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0154] In some embodiments, about 3.5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year. In some cases, about 3.5 g of the anti-C1s antibody is administered to the subject about every twelve weeks for a period of more than 1 year. For example, in some embodiments, about 3.5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0155] In some embodiments, about 3 g to about 5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year, and about 3 g to about 5 g of the antibody is additionally administered on or about day 29. In some cases, about 3 g to about 5 g of the anti-C1s antibody is administered to the subject about every twelve weeks for a period of more than 1 year, and about 3 g to about 5 g of the antibody is additionally administered on or about day 29. For example, in some embodiments, about 3 g to about 5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of 1 year to 50 years, e.g., 1 year to 2 years, 2 years to 5 years, 5 years to 10 years, 10 years to 20 years, 20 years to 30 years, 30 years to 40 years, or 40 years to 50 years, and about 3 g to about 5 g of the antibody is additionally administered on or about day 29.

[0156] In some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of about 12 weeks to 1 year, such as about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered at or about day 29. In some cases, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the anti-C1s antibody is administered to the subject every twelve weeks for a period of more than 1 year, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered at or about day 29. For example, in some embodiments, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of anti-C1s antibody is administered to the subject approximately every 12 weeks for a period of 1 to 50 years, such as 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years, and an additional about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g of the antibody is administered on or about day 29.

[0157] In some embodiments, about 3.5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 24 weeks, about 6 months to about 9 months, or about 6 months to 1 year, and an additional about 3.5 g of the antibody is administered at or about day 29. In some cases, about 3.5 g of the anti-C1s antibody is administered to the subject about every twelve weeks for a period of more than 1 year, and an additional about 3.5 g of the antibody is administered at or about day 29. For example, in some embodiments, about 3.5 g of the anti-C1s antibody is administered to the subject about every 12 weeks for a period of 1 year to 50 years, e.g., 1 year to 2 years, 2 years to 5 years, 5 years to 10 years, 10 years to 20 years, 20 years to 30 years, 30 years to 40 years, or 40 years to 50 years, and an additional about 3.5 g of the antibody is administered at or about day 29.

[0158] In some embodiments, the present disclosure provides a method of treating CAD in a subject in need thereof, the method comprising administering an effective dose of an anti-C1s antibody to the subject, wherein the subject has an antibody plasma concentration of at least about 100 μg / mL, at least about 200 μg / mL, at least about 300 μg / mL, at least about 400 μg / mL, at least about 500 μg / mL, at least about 600 μg / mL, at least about 700 μg / mL, at least about 800 μg / mL, at least about 900 μg / mL, at least about 1000 μg / mL, at least about 1100 μg / mL, at least about 1200 μg / mL, or at least about 1300 μg / mL. The plasma concentration of the anti-C1s antibody in the subject can be measured using techniques known in the art. In some embodiments, the anti-C1s antibody is measured using a direct binding enzyme-linked immunosorbent assay (ELISA). In some embodiments, the anti-C1s antibody is measured using an indirect ELISA. In some embodiments, the anti-C1s antibody is measured using a sandwich ELISA. In some embodiments, anti-C1s antibodies are measured using a competitive ELISA.

[0159] In some embodiments, the disclosure provides a method of increasing the number of reticulocytes in the blood of a subject with CAD, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the blood of the subject by at least about 1.1 times, at least about 1.2 times, at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, at least about 1.6 times, at least about 1.7 times, at least about 1.8 times, at least about 1.9 times, at least about 2.0 times, at least about 2.1 times, at least about 2.2 times, at least about 2.3 times, at least about 2.4 times, at least about 2.5 times, at least about 2.6 times, at least about 2.7 times, at least about 2.8 times, at least about 2.9 times, at least about 3.0 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times after administration.

[0160] In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the subject's blood within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0161] In some embodiments, the present disclosure provides a method of reducing the level of lactate dehydrogenase (LDH) in a subject with CAD, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the anti-C1s antibody reduces the level of LDH to about a normal reference range (about 105-333 IU / L). In some embodiments, the anti-C1s antibody reduces the total level of bilirubin to about 333 IU / L or less.

[0162] In some embodiments, the anti-C1s antibody decreases the level of LDH in the subject within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0163] In some embodiments, the disclosure provides a method of increasing the level of hemoglobin in a subject having CAD, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the anti-C1s antibody increases the level of hemoglobin in the subject by at least about 1.0 g / dL, 1.1 g / dL, 1.2 g / dL, 1.3 g / dL, 1.4 g / dL, 1.5 g / dL, 1.6 g / dL, 1.7 g / dL, 1.8 g / dL, 1.9 g / dL, 2.0 g / dL, 2.1 g / dL, 2.2 g / dL, 2.3 g / dL, 2.4 g / dL, 2.5 g / dL, 2.6 g / dL, 2.7 g / dL, 2.8 g / dL, 2.9 g / dL, 3 g / dL, 4 g / dL, 5 g / dL, 6 g / dL, 7 g / dL, 8 g / dL, 9 g / dL, 10 g / dL, 11 g / dL, 12 g / dL, 13 g / dL, 14 g / dL, 15 g / dL, 16 g / dL, 17 g / dL, 18 g / dL, 19 g / dL, 20 g / dL, 21 g / dL, 22 g / dL, 23 g / dL, 24 g / dL, 25 g / dL L, 2.6g / dL, 2.7g / dL, 2.8g / dL, 2.9g / dL, 3.0g / dL, 3.1g / dL, 3.2g / dL, 3.3g / dL, 3.4g / dL, 3.5g / dL , 3.6g / dL, 3.7g / dL, 3.8g / dL, 3.9g / dL, 4.0g / dL, 4.1g / dL, 4.2g / dL, 4.3g / dL, 4.4g / dL or 4.5g / dL. In some embodiments, the anti-C1s antibody increases the total level of hemoglobin in the subject after administration to at least about 10.0 g / dL, at least about 10.1 g / dL, at least about 10.2 g / dL, at least about 10.3 g / dL, at least about 10.4 g / dL, at least about 10.5 g / dL, at least about 10.6 g / dL, at least about 10.7 g / dL, at least about 10.8 g / dL, at least about 10.9 g / dL , at least about 11.0 g / dL, at least about 11.1 g / dL, at least about 11.2 g / dL, at least about 11.3 g / dL, at least about 11.4 g / dL, at least about 11.5 g / dL, at least about 11.6 g / dL, at least about 11.7 g / dL, at least about 11.8 g / dL, at least about 11.9 g / dL, at least about 12.0 g / dL, at least about 12.1 g / dL, at least about 12.2 g / dL dL, at least about 12.3 g / dL, at least about 12.4 g / dL, at least about 12.5 g / dL, at least about 12.6 g / dL, at least about 12.7 g / dL, at least about 12.8 g / dL, at least about 12.9 g / dL, at least about 13.0 g / dL, at least about 13.1 g / dL, at least about 13.2 g / dL, at least about 13.3 g / dL, at least about 13.4 g / dL, at least about 13.5 g / dL, at least about 13.6 g / dL, at least about 13.7 g / dL, at least about 13.8 g / dL, at least about 13.9 g / dL, at least about 14.0 g / dL, at least about 14.1 g / dL, at least about 14.2 g / dL, at least about 14.3 g / dL, at least about 14.4 g / dL, at least about 14.5 g / dL, at least about 14.6 g / dL, at least about 14.7 g / dL, at least about 14.8 g / dL, at least about 14.9 g / dL, at least about 15.1 g / dL, at least about 15.2 g / dL, at least about 15.3 g / dL, at least about 15.4 g / dL, at least about 15.5 g / dL, at least about 15.6 g / dL, at least about 15.7 g / dL, at least about 15.8 g / dL, at least about 15.9 g / dL, at least about 16.1 g / dL, at least about 16.2 g / dL,8g / dL, at least about 14.9g / dL, at least about 15.0g / dL, at least about 15.1g / dL, at least about 15.2g / dL, at least about 15.3g / dL, at least about 15.4g / dL, at least about 15.5g / dL, at least about 15.6g / dL, at least about 15.7g / dL, at least about 15.8g / dL, at least about 15.9g / dL, at least about 16.0g / dL, at least about 16.1g / dL, at least about 16.2g / dL, at least about 16.3g / dL, at least about 16.4g / dL / dL, at least about 16.5g / dL, at least about 16.6g / dL, at least about 16.7g / dL, at least about 16.8g / dL, at least about 16.9g / dL, at least about 17.0g / dL, at least about 17.1g / dL, at least 17.2g / dL, at least about 17.3g / dL, at least about 17.4g / dL, at least about 17.5g / dL, at least about 17.6g / dL, at least about 17.7g / dL, at least about 17.8g / dL, at least about 17.9g / dL, or at least about 18.0g / dL.

[0164] In some embodiments, the anti-C1s antibody increases the level of hemoglobin in the subject within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0165] In some embodiments, the present disclosure provides a method of reducing the level of bilirubin in a subject with CAD, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the anti-C1s antibody reduces the total level of bilirubin to about a normal reference range (about 1.71 to about 20.5 μmol / L). In some embodiments, the anti-C1s antibody reduces the total level of bilirubin to about 20.5 μmol / L or less.

[0166] In some embodiments, the anti-C1s antibody reduces the total level of bilirubin in the subject within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0167] In some embodiments, the present disclosure provides a method of inhibiting the classical complement pathway in a subject with CAD (e.g., in a fluid, tissue, or organ in the individual) by at least about 75%, at least about 80%, at least about 85%, or at least about 90%, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the classical complement pathway is inhibited by at least about 90%. In some embodiments, the classical complement pathway is inhibited by about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%. The activity level of the classical complement pathway can be determined using any of a variety of methods. As a non-limiting example, the activity of the classical complement pathway can be determined ex vivo, for example, by determining the activity level of the classical complement pathway in a blood, serum, or plasma sample obtained from the individual. For example, the classical complement pathway in a blood, serum, or plasma sample can be activated ex vivo, and the amount of complement component cleavage products (such as C5b-9) generated by such activation can be determined. In some embodiments, the activity level of the classical complement pathway can be determined using a method of inhibiting the classical complement pathway in a subject with CAD. The Classical Pathway Assay of the Complement System determines the activity level of the classical complement pathway.

[0168] In some embodiments, the anti-C1s antibody inhibits the classical complement pathway in the subject within about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0169] In some embodiments, the present disclosure provides a method for reducing the level of 50% hemolytic complement activity (CH50) in a subject with CAD, the method comprising administering an anti-C1s antibody to the subject. The CH50 assay is a standard clinical test for measuring the complement-mediated hemolytic ability of a serum sample. In some embodiments, the CH50 level is reduced by about 10-20 IU / mL, about 10-15 IU / mL, or about 15-20 IU / mL. In some embodiments, the (CH50) level is reduced by about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 IU / mL. In some embodiments, the (CH50) level is reduced to less than about 10 IU / mL, less than about 9 IU / mL, less than about 8 IU / mL, less than about 7 IU / mL, less than about 6 IU / mL, or less than about 5 IU / mL.

[0170] In some embodiments, the anti-C1s antibody decreases CH50 levels in the subject within about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0171] In some embodiments, the inclusion criteria are as follows. Participants are male and female adults ≥ 18 years of age at the time of signing the informed consent and exhibit the following characteristics:

[0172] 1. Diagnosis of CAD is based on the following criteria:

[0173] Chronic hemolysis;

[0174] A positive polyspecific direct antiglobulin test (DAT) status;

[0175] ·Monospecific DAT is strongly positive for C3d;

[0176] Cold agglutinin [CAg] titer ≥ 64 at 4°C;

[0177] IgG DAT ≤1+; and

[0178] No obvious malignant disease

[0179] 2. Hgb levels were ≤10.0 g / dL on days -42 and -21 of the screening period; if the patient received a blood transfusion during the screening period, a qualified Hgb value must be obtained at least 28 days after the transfusion.

[0180] 3. Bilirubin levels above the normal reference range, including participants with Gilbert's syndrome

[0181] 4. Ferritin levels above the lower limit of normal. Concurrent treatment with iron supplements was allowed if the participant had been on a stable dose within the previous 4 weeks.

[0182] 5. Presence of at least moderate CAD-related fatigue based on the "Fatigue Screener" conducted on Screening Period Day -42. That is, participants must report having moderate fatigue, severe fatigue, or very severe fatigue related to CAD in the past month.

[0183] In some embodiments, the methods of the present disclosure prevent or delay the onset of at least one symptom of CAD in a subject. In some embodiments, the compositions of the present disclosure reduce or eliminate at least one symptom of CAD in a subject. The symptom may also be the activity of C1s protein in the cells, tissues, or fluids of the individual. The symptom may also be the degree of complement activation in the cells, tissues, or fluids of the individual.

[0184] "Treating" means at least ameliorating a symptom associated with CAD, wherein ameliorating is used in a broad sense to mean at least reducing the magnitude of a parameter (eg, symptom) associated with CAD.

[0185] Baseline values ​​for laboratory parameters related to anemia (Hgb) and hemolysis (bilirubin) and baseline values ​​related to fatigue (FACIT-fatigue scale score) were defined as the average of the assessments on Day -21 during the screening period and on Day 1 before treatment administration. If a transfusion occurred during the screening period, the baseline measurement must be at least 28 days after the transfusion. In that case, any measurement within 28 days of the transfusion was considered invalid and was subsequently excluded, and the baseline value was then the last valid measurement before treatment administration.

[0186] The primary treatment assessment of anemia-related laboratory parameters was the mean of nonmissing measurements of Hgb assessed on Days 141, 155, and 169 [Weeks 21, 23, and 25].

[0187] Endpoint responder rate: The percentage of participants who achieved an increase of ≥2 g / dL from baseline based on the mean of Hgb values ​​measured on Days 141, 155, and 169 [Weeks 21, 23, and 25] and who met both of these criteria: (1) no transfusions during the last 21 weeks; and (2) no use of protocol-prohibited CAD medications from baseline.

[0188] The primary treatment assessment measured by the FACIT-Fatigue scale score was the mean of the non-missing FACIT-Fatigue scale scores assessed at the same time points on Days 141, 155, and 169. Endpoint: Change from baseline in the FACIT-Fatigue measure at the primary treatment assessment time point.

[0189] There are three key secondary endpoints: 1. Change from baseline in bilirubin at the primary treatment assessment time point; 2. Change from baseline in Hgb at Day 29; and 3. Change from baseline in FACIT-Fatigue scale score at Day 29. Another secondary endpoint is: mean change from baseline in bilirubin at Day 29.

[0190] A variety of hosts (wherein the term "host" is used interchangeably with the terms "subject", "individual" and "patient" herein) can be treated according to the subject methods. Typically, such hosts are "mammals" or "mammals", wherein these terms are widely used to describe organisms within the class Mammalia, including carnivores (e.g., cats), herbivores (e.g., cattle, horses and sheep), omnivores (e.g., dogs, goats and pigs), rodents (e.g., mice, guinea pigs and rats) and primates (e.g., humans, chimpanzees and monkeys). In some embodiments, the host is an individual with a complement system, such as a mammal, fish or invertebrate. In some embodiments, the host is a mammal, fish or invertebrate companion animal, agricultural animal, working animal, zoo animal or laboratory animal containing a complement system. In some embodiments, the host is a human.

[0191] Treatment options for CIDP

[0192] Chronic inflammatory demyelinating polyneuropathy (CIDP) is a rare immune-mediated disorder of the peripheral nervous system characterized by loss of myelin in motor and sensory nerves, manifested by weakness, hyporeflexia or areflexia, numbness, paresthesias, and sensory ataxia, indicative of large sensory fiber involvement.

[0193] The diagnosis of CIDP is usually confirmed by neurophysiological evidence of peripheral nerve demyelination and exclusion of other causes of demyelinating polyneuropathy or diseases that may have similar presentations. Common expert consensus criteria for diagnosing CIDP have been provided by the Joint Task Force of the European Federation of Neurological Societies / Peripheral Nerve Society (EFNS / PNS) (2010) (Bergh Van den PYK. et al. European J. of Neurology [European Journal of Neurology] 2010, 17: 356-363), and recent revisions of these criteria (2021) are available (Van den Bergh et al. Eur J Neurol [European Journal of Neurology] 2021; 28(11): 3556-3583).

[0194] Activation of the complement pathway is expected to play a key role in the pathophysiology of CIDP. In CIDP patients, this pathway (enzymatic cascade) is triggered by pathogenic autoantibodies, causing inflammatory damage, ultimately driving a demyelinating and inflammatory process in peripheral nerves that underlies functional disability.

[0195] By more selectively targeting key mechanisms of the disease, the anti-C1s antibodies of the present disclosure have the potential to improve efficacy profiles in a broad CIDP population, including patients refractory to standard of care (SOC) therapies.

[0196] The present disclosure provides methods for treating CIDP in a subject. CIDP can be typical or atypical (Doneddu et al., Curr Opin Neurol. 34(5):613-624 (2021). In some embodiments, CIDP can be typical CIDP. In some embodiments, CIDP can be atypical CIDP. In some embodiments, atypical CIDP can exist together with Lewis-Sumner syndrome (LSS). In some embodiments, atypical CIDP can exist together with pure motor CIDP. In one aspect, these methods include administering a loading dose of an anti-C1s antibody to the subject, followed by one or more maintenance doses of the antibody.

[0197] In some embodiments, the loading dose is a body weight based dose. In some embodiments, the loading dose of the antibody is about 50 mg / kg body weight of the subject.

[0198] In some embodiments, the maintenance dose of the antibody is a fixed dose. In some embodiments, the maintenance dose of the antibody is between about 200mg to about 800mg. In some embodiments, the maintenance dose of the antibody is between about 200mg to about 300mg, about 300mg to about 400mg, about 200mg to about 500mg, about 300mg to about 600mg, about 400mg to about 800mg or about 600mg to about 800mg. In some embodiments, the maintenance dose of the antibody is between about 300mg to about 600mg. In some embodiments, the maintenance dose of the antibody is about 300mg, about 400mg, about 500mg or about 600mg. In some embodiments, the maintenance dose of the antibody is about 300mg. In some embodiments, the maintenance dose of the antibody is about 600mg.

[0199] In some embodiments, the maintenance dose of the antibody is a fixed dose. In some embodiments, the maintenance dose of the antibody is between about 200mg to about 800mg. In some embodiments, the maintenance dose of the antibody is between about 200mg to about 300mg, about 300mg to about 400mg, about 200mg to about 500mg, about 300mg to about 600mg, about 400mg to about 800mg or about 600mg to about 800mg. In some embodiments, the maintenance dose of the antibody is between about 300mg to about 600mg. In some embodiments, the maintenance dose of the antibody is about 300mg, about 400mg, about 500mg or about 600mg. In some embodiments, the maintenance dose of the antibody is about 300mg. In some embodiments, the maintenance dose of the antibody is about 600mg.

[0200] In some embodiments, one or more maintenance doses are administered about weekly (ie, every week), every 2 weeks, every 4 weeks, or every 12 weeks.

[0201] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose approximately weekly starting on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose approximately weekly starting on day 8, subcutaneously.

[0202] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of between about 200 mg to about 800 mg approximately weekly beginning on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of between about 200 mg to about 800 mg approximately weekly beginning on day 8, subcutaneously.

[0203] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of between about 1000 mg to about 8000 mg about every 2 weeks, about every 4 weeks, or about every 12 weeks starting on day 8 or day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of between about 1000 mg to about 8000 mg about every 2 weeks, about every 4 weeks, or about every 12 weeks starting on day 8 or day 29.

[0204] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of between about 300 mg to about 600 mg approximately weekly beginning on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of between about 300 mg to about 600 mg approximately weekly beginning on day 8, subcutaneously.

[0205] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of between about 1200 mg to about 7200 mg about every 4 weeks or about every 12 weeks starting on day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of between about 1200 mg to about 7200 mg about every 4 weeks or about every 12 weeks starting on day 29.

[0206] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 300 mg approximately weekly beginning on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 300 mg approximately weekly beginning on day 8, subcutaneously.

[0207] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 600 mg approximately weekly beginning on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 600 mg approximately weekly beginning on day 8, subcutaneously.

[0208] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 600 mg approximately every 2 weeks starting on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 600 mg approximately every 2 weeks starting on day 8, subcutaneously.

[0209] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 1200 mg approximately every 2 weeks starting on day 8. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 1200 mg approximately every 2 weeks starting on day 8 subcutaneously.

[0210] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 1200 mg approximately every 4 weeks starting on day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 1200 mg approximately every 4 weeks starting on day 29.

[0211] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 2400 mg approximately every 4 weeks starting on day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 2400 mg approximately every 4 weeks starting on day 29.

[0212] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 3600 mg approximately every 12 weeks starting on day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 3600 mg approximately every 12 weeks starting on day 29.

[0213] In some embodiments, a loading dose is administered on day 1, followed by a maintenance dose of about 7200 mg approximately every 12 weeks starting on day 29. In some embodiments, a loading dose is administered intravenously on day 1, followed by a maintenance dose of about 7200 mg subcutaneously every 12 weeks starting on day 29.

[0214] In some embodiments, a maintenance dose of between about 200 mg and about 800 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of between about 200 mg and about 800 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of more than 1 year. For example, in some embodiments, a maintenance dose of between about 200 mg and about 800 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0215] In some embodiments, a maintenance dose of between about 1000 mg and about 8000 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or every 4 weeks, or about every 12 weeks, for a period of about 4 weeks to 1 year, such as about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of between about 1000 mg and about 8000 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or about every 4 weeks, or about every 12 weeks, for a period of more than 1 year. For example, in some embodiments, a maintenance dose of between about 1000 mg and about 8000 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or about every 4 weeks, or about every 12 weeks, for a period of 1 to 50 years, such as 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0216] In some embodiments, a maintenance dose of between about 300 mg to about 600 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of between about 300 mg to about 600 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of more than 1 year. For example, in some embodiments, a maintenance dose of between about 300 mg to about 600 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0217] In some embodiments, a maintenance dose of between about 1200 mg and about 7200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or about every 4 weeks, or about every 12 weeks, for a period of about 4 weeks to 1 year, such as about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of between about 1200 mg and about 7200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or about every 4 weeks, or about every 12 weeks, for a period of more than 1 year. For example, in some embodiments, a maintenance dose of between about 1200 mg and about 7200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks, or about every 4 weeks, or about every 12 weeks, for a period of 1 to 50 years, such as 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0218] In some embodiments, a maintenance dose of about 300 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 300 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 300 mg of an anti-C1s antibody is administered to a subject approximately weekly for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0219] In some embodiments, a maintenance dose of about 600 mg of an anti-C1s antibody is administered to a subject about weekly or about every 2 weeks for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 600 mg of an anti-C1s antibody is administered to a subject about weekly or about every 2 weeks for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 600 mg of an anti-C1s antibody is administered to a subject about weekly or about every 2 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0220] In some embodiments, a maintenance dose of about 1200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks or about every 4 weeks for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 1200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks or about every 4 weeks for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 1200 mg of an anti-C1s antibody is administered to a subject about every 2 weeks or about every 4 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0221] In some embodiments, a maintenance dose of about 2400 mg of an anti-C1s antibody is administered to a subject about every 4 weeks for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 2400 mg of an anti-C1s antibody is administered to a subject about every 4 weeks for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 2400 mg of an anti-C1s antibody is administered to a subject about every 4 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0222] In some embodiments, a maintenance dose of about 3,600 mg of an anti-C1s antibody is administered to a subject about every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 3,600 mg of an anti-C1s antibody is administered to a subject about every 12 weeks for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 3,600 mg of an anti-C1s antibody is administered to a subject about every 12 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0223] In some embodiments, a maintenance dose of about 7200 mg of an anti-C1s antibody is administered to a subject approximately every 12 weeks for a period of about 12 weeks to 1 year, e.g., about 12 weeks to about 8 weeks, about 2 months to about 6 months, about 6 months to about 9 months, or about 6 months to 1 year. In some embodiments, a maintenance dose of about 7200 mg of an anti-C1s antibody is administered to a subject approximately every 12 weeks for a period of more than 1 year. For example, in some embodiments, a maintenance dose of about 7200 mg of an anti-C1s antibody is administered to a subject approximately every 12 weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0224] In some embodiments, a one-time 50 mg / kg intravenous loading dose is administered on day 1, and a subcutaneous (SC) dose is administered with a syringe, or a prefilled syringe, or an automatic injector, or a large volume drug delivery system. In some embodiments, the SC dose administered is 300 mg qw, 600 mg qw, 600 mg q2w, 1200 mg q2w, 1200 mg q4w, 2400 mg q4w, 3600 mg q12w, or 7200 mg q12w. In some embodiments, SC doses are administered with a syringe, a prefilled syringe, or an automatic injector, or a large volume drug delivery system until the end of treatment.

[0225] In some embodiments, the subject is refractory to another CIDP treatment, also referred to herein as "standard of care (SOC) refractory." SOC refractory patients are patients with evidence of failure or inadequate response to SOC, defined as no clinically meaningful improvement after a minimum of 12 weeks of SOC treatment and a persistent Inflammatory Neuropathy Causes and Treatment (INCAT) score of ≥ 2 (Hughes et al. Ann Neurol. 2001; 50(2): 195-201), or patients who are unable to receive or continue immunoglobulin or corticosteroid therapy due to side effects. In some embodiments, clinically meaningful improvement (and deterioration) is defined as one of the following: a decrease (increase) of ≥1 point in the adjusted INCAT score, an increase (decrease) of ≥4 points in the Inflammatory Rasch Global Disability Scale (I-RODS) total score (van Nes et al. Neurology. 2011; 76(4):337-45), an increase (decrease) of ≥3 points in the Medical Regulatory Committee (MRC) total score, an improvement (decrease) of ≥8 kPa in the mean grip strength (one hand), or an equivalent improvement (deterioration) based on information recorded in the medical record.

[0226] In some embodiments, the subject is being treated with another CIDP treatment, also referred to herein as "SOC treated". In some embodiments, the subject receives another CIDP treatment within about one week of the loading dose, or receives another CIDP treatment simultaneously. Patients treated with SOC are patients receiving stable SOC therapy who show documented evidence of objective and clinically significant responses, as well as evidence of clinically significant deterioration when SOC therapy is interrupted or the dose is reduced. In some embodiments, clinically significant improvement (and deterioration) is defined as one of the following: a decrease (increase) of ≥1 point in the adjusted INCAT score, an increase (decrease) of ≥4 points in the I-RODS total score, an increase (decrease) of ≥3 points in the MRC total score, an improvement (decrease) of ≥8 kPa in the average grip strength (one hand), or an equivalent improvement (deterioration) based on information recorded in the medical record.

[0227] In some embodiments, the subject is not treated for CIDP, also referred to herein as "not treated for SOC". Patients who are not treated for SOC are patients who have not previously been treated for CIDP or who have received immunoglobulin (IVIg or SCIg) or corticosteroids but have stopped treatment for reasons other than lack of response or side effects. Patients should not have received immunoglobulin (IVIg or SCIg) or corticosteroid treatment within at least 6 months prior to screening.

[0228] Other CIDP treatments can be any known CIDP treatments. In some embodiments, other CIDP treatments are intravenous immunoglobulin (IVIg), subcutaneous immunoglobulin (SCIg), corticosteroids (e.g., prednisone, prednisolone, methylprednisolone), cyclophosphamide, rituximab, tocilizumab, azathioprine, tacrolimus, plasma exchange, mycophenolate mofetil, cyclosporine, plasma exchange, or a combination thereof. Corticosteroid treatment can be oral or intravenous.

[0229] In some embodiments, the administration of an anti-C1s antibody improves the adjusted INCAT disability score. In some embodiments, the administration of an anti-C1s antibody reduces the INCAT score by 1 point or more. In some embodiments, the subject is SOC refractory. In some embodiments, the subject is SOC naive. The INCAT disability scale is an effective tool for measuring disability in CIDP patients. It has been used in large clinical trials leading to the approval of IVIg and SCIg (PATH). The total INCAT score is obtained by adding the scores corresponding to the arms and legs.

[0230] In some embodiments, the primary endpoint is the percentage of participants with an adjusted INCAT score decrease of 1 point or more from baseline (responder) at Week 24. The adjusted INCAT disability score is the same as the INCAT disability score but excludes changes in upper limb function from 0 (normal) to 1 (mild symptoms or signs in 1 or both arms that do not affect any function listed on the scale) or from 1 to 0.

[0231] In some embodiments, inclusion and exclusion criteria are as follows. Participants are male and female adults (≥18 years old) with CIDP or probable CIDP criteria based on the EAN / PNS Working Group CIDP Guidelines, 2nd Revision (2021). Participants with CIDP must have classic CIDP, pure motor CIDP, or Lewis-Summer syndrome, and no Other polyneuropathies, including pure sensory CIDP or distal CIDP.

[0232] Participants must also:

[0233] i. Have responded to IVIg in the past. The response must be an objective, clinically meaningful improvement defined by at least one of the following: a decrease of ≥1 point in the adjusted INCAT score, an increase of ≥4 points in the I-RODS total score, an increase of ≥3 points in the MRC total score, an improvement of ≥8 kPa in mean grip strength (one hand), or an equivalent improvement based on information documented in the medical record and at the discretion of the PI.

[0234] ii. Receive IVIg within 20% of the standard maintenance dosing regimen. The standard maintenance dose is defined as 0.4-1 g / kg every 2-6 weeks (EAN / PNS2021 guidelines).

[0235] iii. Receiving a stable maintenance dose of IVIg, defined as no more than a 10% change in the frequency or dose of IVIg in the 8 weeks prior to screening and remaining stable until baseline.

[0236] iv. Have residual disability, defined as an adjusted INCAT score of 2-9 at screening confirmed at baseline.

[0237] v.Previous vaccination against enveloped organisms.

[0238] In some embodiments, administration of an anti-C1s antibody prevents relapse. Relapse is defined as an increase of 1 or more in the adjusted INCAT disability score. In some embodiments, administration of an anti-C1s antibody prevents relapse after withdrawal of another CIDP treatment.

[0239] In some embodiments, administration of an anti-C1s antibody improves the MRC total score by 3 points or more.

[0240] In some embodiments, administration of the anti-C1s antibody improves the I-RODS score by 4 points or more.

[0241] In some embodiments, administration of an anti-C1s antibody improves average grip strength (eg, one hand) by 8 kilopascals or more.

[0242] In some embodiments, the present disclosure provides a method of treating CIDP in a subject in need thereof, the method comprising administering an effective dose of an anti-C1s antibody to the subject, wherein the subject has an antibody plasma concentration of at least about 100 μg / mL, at least about 200 μg / mL, at least about 300 μg / mL, at least about 400 μg / mL, at least about 500 μg / mL, at least about 600 μg / mL, at least about 700 μg / mL, at least about 800 μg / mL, at least about 900 μg / mL, at least about 1000 μg / mL, at least about 1100 μg / mL, at least about 1200 μg / mL, at least about 1300 μg / mL, at least about 1400 μg / mL, or at least about 1500 μg / mL. The plasma concentration of the anti-C1s antibody in the subject can be measured using techniques known in the art. In some embodiments, the anti-C1s antibody is measured using a direct binding enzyme-linked immunosorbent assay (ELISA). In some embodiments, the anti-C1s antibody is measured using an indirect ELISA. In some embodiments, anti-C1s antibodies are measured using a sandwich ELISA. In some embodiments, anti-C1s antibodies are measured using a competitive ELISA.

[0243] In some embodiments, the present disclosure provides a method of inhibiting the classical complement pathway in a subject with CIDP (e.g., in a fluid, tissue, or organ in the individual) by at least about 75%, at least about 80%, at least about 85%, or at least about 90%, the method comprising administering an anti-C1s antibody to the subject. In some embodiments, the classical complement pathway is inhibited by at least about 90%. In some embodiments, the classical complement pathway is inhibited by about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%. The activity level of the classical complement pathway can be determined using any of a variety of methods. As a non-limiting example, the activity of the classical complement pathway can be determined ex vivo, for example, by determining the activity level of the classical complement pathway in a blood, serum, or plasma sample obtained from the individual. For example, the classical complement pathway in a blood, serum, or plasma sample can be activated ex vivo, and the amount of complement component cleavage products (such as C5b-9) generated by such activation can be determined. In some embodiments, the activity level of the classical complement pathway can be determined using a method of inhibiting the classical complement pathway in a subject with CIDP. The Classical Pathway Assay of the Complement System determines the activity level of the classical complement pathway.

[0244] In some embodiments, the anti-C1s antibody inhibits the classical complement pathway in the subject within about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0245] In some embodiments, the present disclosure provides a method for reducing the level of 50% hemolytic complement activity (CH50) in a subject with CIDP, the method comprising administering an anti-C1s antibody to the subject. The CH50 assay is a standard clinical test for measuring the complement-mediated hemolytic ability of a serum sample. In some embodiments, the CH50 level is reduced by about 10-20 IU / mL, about 10-15 IU / mL, or about 15-20 IU / mL. In some embodiments, the (CH50) level is reduced by about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 IU / mL. In some embodiments, the (CH50) level is reduced to less than about 10 IU / mL, less than about 9 IU / mL, less than about 8 IU / mL, less than about 7 IU / mL, less than about 6 IU / mL, or less than about 5 IU / mL.

[0246] In some embodiments, the anti-C1s antibody decreases CH50 levels in the subject within about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, or about 12 weeks of administration.

[0247] In some embodiments, the methods of the present disclosure prevent or delay the onset of at least one symptom of CIDP in a subject. In some embodiments, the compositions of the present disclosure reduce or eliminate at least one symptom of CIDP in a subject. The symptom may also be the activity of C1s protein in the cells, tissues, or fluids of the individual. The symptom may also be the degree of complement activation in the cells, tissues, or fluids of the individual.

[0248] "Treatment" means at least ameliorating the symptoms associated with CIDP, wherein ameliorating is used in a broad sense to mean at least reducing the magnitude of a parameter (eg, symptom) associated with CIDP.

[0249] A variety of hosts (wherein the term "host" is used interchangeably with the terms "subject", "individual" and "patient" herein) can be treated according to the subject methods. Typically, such hosts are "mammals" or "mammals", wherein these terms are widely used to describe organisms within the class Mammalia, including carnivores (e.g., cats), herbivores (e.g., cattle, horses and sheep), omnivores (e.g., dogs, goats and pigs), rodents (e.g., mice, guinea pigs and rats) and primates (e.g., humans, chimpanzees and monkeys). In some embodiments, the host is an individual with a complement system, such as a mammal, fish or invertebrate. In some embodiments, the host is a mammal, fish or invertebrate companion animal, agricultural animal, working animal, zoo animal or laboratory animal containing a complement system. In some embodiments, the host is a human.

[0250] Composition

[0251] The anti-C1s antibody is generally present in a composition (eg, a pharmaceutical composition). The humanized anti-C1s antibody of the present disclosure can be formulated into a pharmaceutical composition by combining with an appropriate pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent or other pharmaceutically acceptable excipient.

[0252] Exemplary antibody concentration ranges in the compositions of the present disclosure may be about 50 mg / mL to about 250 mg / mL, about 75 mg / mL to about 225 mg / mL, about 100 mg / mL to about 200 mg / mL, about 100 mg / mL to about 150 mg / mL, about 150 mg / mL to about 200 mg / mL, about 125 mg / mL to about 175 mg / mL, or about 140 mg / mL to about 160 mg / mL. In some embodiments, the antibody concentration in the compositions of the present disclosure is about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, about 130 mg / mL, about 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 175 mg / mL, about 200 mg / mL, or about 250 mg / mL. In some embodiments, the compositions of the present disclosure comprise about 150 mg / mL of humanized anti-C1s antibody.

[0253] In some embodiments, a composition comprising an anti-C1s antibody comprises one or more of a stabilizer, a buffer, a cryoprotectant, a chelating agent, and a surfactant.

[0254] Suitable stabilizers include arginine or a salt thereof (e.g., arginine citrate, arginine hydrochloride, arginine oxalate, arginine phosphate, arginine succinate or arginine tartrate), sugars, sugar alcohols or amino sugars (e.g., fructose, galactose, glucose, lactose, maltose, mannose, raffinose, sorbitol, sorbose, sucrose, galactosamine, glucosamine, N-methylglucosamine and neuraminic acid). Other stabilizers may also be used.

[0255] Suitable buffers include acetate, citrate, histidine, oxalate, phosphate, succinate, and tartrate. Other buffers may also be used.

[0256] Suitable cryoprotectants include sucrose, ethylene glycol, dimethyl sulfoxide (DMSO), glycerol, trehalose, and propylene glycol. Other cryoprotectants may also be used.

[0257] Suitable chelating agents include ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetate (DTPA), dihydroxyethylglycine, citric acid, tartaric acid and methionine. Other chelating agents may also be used.

[0258] Suitable surfactants include polysorbates (e.g., polysorbate 20 (PS20), polysorbate 40 (PS40), polysorbate 60 (PS60), and polysorbate 80 (PS80)), dicarboxylic acids, oxalic acid, succinic acid, fumaric acid, phthalic acid, polyoxyethylene sorbitan monooleate, poloxamers (e.g., P188), and polyethylene glycol. Other surfactants may also be used.

[0259] The compositions of the present disclosure may have a pH of about 6 to about 7.5, about 6 to about 7, about 6.5 to about 7.5, or about 6.5 to about 7.1. In some embodiments, the compositions of the present disclosure have a pH of about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, or about 7.5. In some embodiments, the compositions of the present disclosure have a pH of about 6.5-7.1. In some embodiments, the compositions of the present disclosure have a pH of about 6.8.

[0260] The compositions of the present disclosure may contain other excipients, including but not limited to water for injection, diluents, solubilizers, soothing agents, additional buffers, inorganic or organic salts, antioxidants, etc. Pharmaceutically acceptable excipients are readily available to the public. In some embodiments, the compositions of the present disclosure do not contain other excipients other than those described above. Other pharmaceutically acceptable carriers or excipients (such as Remington's Pharmaceutical Sciences 16 th edition [Remington's Pharmaceutical Sciences, 16th Edition], Osol, A. ed. (1980)) can be included in the formulation, provided that they do not adversely affect the desired properties of the formulation. In some embodiments, a preservative can be added. In some embodiments, the composition is substantially free of preservatives. Cryoprotectants or lyoprotectants can be included in the lyophilized formulation.

[0261] The compositions of the present disclosure may be in liquid form, lyophilized form (wherein the lyophilized formulation is reconstituted with a sterile solution prior to administration), or in liquid form reconstituted from a lyophilized form. The standard procedure for reconstructing a lyophilized composition is to add back a certain volume of pure water (usually equivalent to the volume removed during lyophilization); however, a solution containing an antibacterial agent can be used to produce a pharmaceutical composition for parenteral administration; see also Chen (1992) Drug Dev Ind Pharm [Drug Development and Industrial Pharmaceuticals] 18, 1311-54. In some embodiments, the compositions of the present disclosure are in liquid form. The liquid formulation can be prepared and used immediately, or it can be diluted before injection.

[0262] Route of administration

[0263] The antibodies of the present disclosure may be administered to a subject using any available conventional methods and routes suitable for delivering conventional drugs, including systemic or local routes. Generally, routes of administration contemplated by the present disclosure include, but are not necessarily limited to, enteral, parenteral or inhalation routes.

[0264] Parenteral administration routes other than administration by inhalation include, but are not necessarily limited to, topical, transdermal, subcutaneous, intramuscular, intraorbital, intracapsular, intraspinal, intrasternal, intrathecal, and intravenous routes, i.e., any route of administration other than through the digestive tract. Parenteral administration can be performed to achieve systemic or local delivery of the subject antibody. Where systemic delivery is desired, administration typically involves local or mucosal administration of the invasive or systemic absorption of the pharmaceutical formulation.

[0265] If desired, the administration routes can be combined or adjusted according to the desired effect. The antibody can be administered in a single dose or multiple doses. In some embodiments, the antibodies of the present disclosure are administered intravenously. In some embodiments, the antibodies of the present disclosure are administered subcutaneously.

[0266] The present disclosure also provides a pharmaceutical unit dosage form comprising a therapeutically effective amount of the composition of the present disclosure for treating one or more complement-mediated diseases in a subject by administering the dosage form to the subject. In some embodiments, the subject is a human. The term "pharmaceutical unit dosage form" refers to physically discrete units suitable as unit doses for subjects to be treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic / preventive effect.

[0267] The unit dosage form can be a container containing the preparation. Suitable containers include, but are not limited to, sealed ampoules, vials (e.g., glass vials), bottles, syringes, and test tubes. The container can be formed of various materials (such as glass or plastic) and can have a sterile access port (e.g., the container can be a vial with a stopper that can be pierced by a hypodermic needle). In some embodiments, the container is a vial. In some embodiments, the container is a prefilled syringe. Typically, the container should maintain the sterility and stability of the preparation.

[0268] An exemplary drug delivery device may involve a needle-based injection system as described in Table 1 of Section 5.2 of ISO 11608-1:2022. As described in ISO 11608-1:2022, needle-based injection systems can be broadly divided into multi-dose container systems and single-dose (partially or completely emptied) container systems. The container may be a replaceable container or an integral non-replaceable container.

[0269] As further described in ISO 11608-1:2022, a multiple-dose container system may involve a needle-based injection device with a replaceable container. In such a system, each container holds multiple doses, the sizes of which may be fixed or variable (pre-set by the user). Another multiple-dose container system may involve a needle-based injection device with an integral non-replaceable container. In such a system, each container holds multiple doses, the sizes of which may be fixed or variable (pre-set by the user).

[0270] As further described in ISO 11608-1:2022, a single dose container system can relate to a needle-based injection device with a replaceable container. In one example of such a system, each container holds a single dose, wherein the entire deliverable volume is discharged (completely emptied). In another example, each container holds a single dose, wherein a portion of the deliverable volume is discharged (partially emptied). Also as described in ISO 11608-1:2022, a single dose container system can relate to a needle-based injection device with an integral non-replaceable container. In one example of such a system, each container holds a single dose, wherein the entire deliverable volume is discharged (completely emptied). In another example, each container holds a single dose, wherein a portion of the deliverable volume is discharged (partially emptied).

[0271] An exemplary sleeve-triggered autoinjector with a manual needle inserter is described in International Publication WO 2015 / 004052. Example audible end-of-dose feedback mechanisms are described in International Publications WO 2016 / 193346 and WO 2016 / 193348. An example needle safety mechanism after use of an autoinjector is described in International Publication WO 2016 / 193352. An example needle sheath remover mechanism for a syringe autoinjector is described in International Publication WO 2016 / 193353. An example support mechanism for supporting the axial position of a syringe is described in International Publication WO 2016 / 193355.

[0272] Kits and Products

[0273] The present disclosure provides a kit or article comprising the composition of the present disclosure. The kit or article comprises a container containing the antibody of the present disclosure. The kit or article may further comprise one or more containers comprising a pharmaceutically acceptable excipient, and include other materials desired from a commercial and user perspective, including filters, needles and syringes. Associated with the kit may be instructions, which are generally included in the commercial packaging of the treatment, prevention or diagnosis product, including information about, for example, the indications, usage, dosage, manufacture, administration, contraindications and / or warnings for the use of such treatment, prevention or diagnosis products. The kit may also be associated with a label, which may be any type of data carrier (e.g., leaflet, sticker, chip, printed matter or barcode) containing information. In certain embodiments, the instructions as listed above may be included in or on a label. The kit may further comprise a device for administering a formulation, and in particular a device containing a composition, i.e. a pre-filled device, such as, but not limited to, a pre-filled syringe or a pre-filled automatic syringe. The kit may also comprise a container containing an antibody, i.e., a pre-filled container, such as a pre-filled vial, cartridge, pouch or ampoule.

[0274] Examples

[0275] Example 1. First-in-human study of SAR445088

[0276] SAR445088 is a humanized monoclonal antibody that selectively inhibits activated C1s and prevents the enzymatic action of C1 on its substrates C4 and C2, thereby inhibiting the formation of the classical pathway C3 convertase C4b2a. SAR445088 inhibits the downstream signaling cascade of the complement system originating via the CP, while the alternative pathway (AP) and the lectin pathway remain functional.

[0277] SAR445088 contains a serine-proline mutation (S241P) in the core-hinge region of the stabilizing molecule, a leucine to glutamate mutation (L248E) that eliminates Fcγ receptor binding, and a methionine to leucine mutation (M428L) and an asparagine to serine mutation (N434S) that increases neonatal Fc receptor (FcRn) binding affinity and increases half-life. SAR445088's LC CDR1, LC CDR2, LC CDR3, HC CDR1, HC CDR2, HC CDR3, VH, VL, and heavy and light chain sequences are described elsewhere in this application as SEQ ID NO.1-10. The sequence is also described in U.S. Patent No. 9,512,233, U.S. Patent No. 10,729,767, and U.S. Patent Application Publication No. US 2020 / 0048332, each of which is incorporated herein by reference in its entirety.

[0278] In nonclinical studies, SAR445088 showed a graded inhibition-concentration relationship.This first-in-human (FIH) clinical trial investigated the safety, tolerability, pharmacokinetic (PK) and pharmacodynamic (PD) profiles of single intravenous (IV) and subcutaneous (SC) administration and multiple SC doses of SAR445088 in healthy participants.

[0279] method

[0280] Study Design

[0281] The FIH trial is a single-center, double-blind, randomized, placebo-controlled, comprehensive single ascending dose (SAD; Part 1) and multiple ascending dose (MAD; Part 2) study of SAR445088 in healthy participants, and includes a Japanese cohort. In Part 1, nine dose cohorts (IV: 2, 4, 8, 16, 30 and 50 mg / kg; SC: 300, 600 and 1200 mg) were planned, with eight subjects per dose cohort, and SAR445088 (n = 6) and matching placebo (n = 2) were randomly assigned at a ratio of 3: 1. For Part 2, three cohorts were planned, with 10 participants per cohort, and SAR445088 (n = 8) and matching placebo (n = 2) were randomly assigned at a ratio of 4: 1. Participants in Part 2 received SAR445088 at 300 or 600 mg SC (once a week for 4 weeks) or 600 mg SC (once a day for 3 consecutive days) or placebo.

[0282] Healthy male and female participants (age: 18–50 years, weight: 45.0–110.0 kg, and body mass index: 18–32 kg / m 2 During the screening period, vaccinations against encapsulated bacterial pathogens, such as meningococci, were administered to participants who had not received these vaccinations within 5 years prior to recruitment.

[0283] Research objectives

[0284] The primary objective of this study is to evaluate the safety and tolerability of SAR445088 in healthy participants. Secondary objectives include evaluation of PK and PD profiles and immunogenicity.

[0285] Safety Assessment

[0286] Safety assessments included physical examination, clinical laboratory tests, serological tests (including a SLE panel), urinalysis, vital signs, anti-drug antibodies (ADA), adverse events (AEs), and electrocardiogram (ECG).

[0287] Bioanalytical methods

[0288] Quantification of SAR445088: SAR445088 in plasma was measured using a high performance liquid chromatography assay with tandem mass spectrometry (MS / MS) detection. SAR445088 was enriched from human plasma using magnetic beads coated with protein G using an immunoaffinity approach. Following standard protein denaturation, reduction, and alkylation treatment steps, the bound protein was subjected to on-bead proteolysis with trypsin. As a result of the trypsin digestion, characteristic peptide fragments derived from the surrogate were used to determine the SAR445088 concentration. The lower limit of quantification (LLOQ) was 10 μg / mL.

[0289] Wieslab CP assay: Quantitative determination of complement activation in serum using an enzyme-linked immunosorbent assay (ELISA). The LLOQ is 10% activity.

[0290] Complement-mediated hemolytic capacity (CH50) assay: CH50 was measured using an in vitro automated homogenized liposome-based assay (Fujifilm Wako) for the quantitative determination of total complement activity in human serum. The LLOQ of this assay was 10 IU / mL.

[0291] Estimation of anti-drug antibodies (ADA): The presence of ADA in plasma was evaluated using an electrochemiluminescent immunoassay (PPD Laboratories, Richmond, VA).

[0292] Estimation of C4: C4 was quantitatively determined using a commercially available in vitro diagnostic (Beckman Coulter, Brea, CA) assay kit from Gen X Laboratories, Inc. (Los Angeles, CA). The LLOQ of this assay was 8 mg / dL.

[0293] Wieslab Alternative Pathway (AP) Assay: An ELISA assay was used to qualitatively determine functional complement AP in human serum with an LLOQ of 10%.

[0294] Pharmacokinetic evaluation: PK parameters were calculated by non-compartmental analysis. SAR445088 plasma concentrations and PK for each dose level and each route (IV and SC) were summarized by descriptive statistics (such as mean, geometric mean, median, standard deviation [SD], standard error of the mean (SEM) and coefficient of variation (CV), minimum and maximum) using PKDMS (in-house software) version 3.1 and Phoenix WinNonLin Professional version 8.1. The data were also summarized by ADA status. For each statement, the mean concentration and PK parameter values ​​are arithmetic means unless otherwise stated. When calculating the mean, concentration values ​​below the plasma assay limit were considered zero. At the time of reaching the maximum drug concentration (t max ), data below the limit of quantitation (BLQ) were considered missing / omitted. The mean concentration value BLQ is reported as BLQ in the table and if the concentration was not observed at the time (t last ) are not drawn in the figure.

[0295] Population PK / PD Modeling: Population PK analysis was performed using data from Parts 1 and 2 using nonlinear mixed effects modeling (NONMEM; Version 7.4.1). Graphical screening of covariates including demographics (sex, age, weight, and race), ADA status, baseline complement factor C4, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) was performed. Statistical evaluation of the selected covariates (ADA status, sex, age, weight, and race) was performed using a stepwise forward selection / backward elimination step. The relationship between SAR445088 exposure and PD response was examined. The effects of covariates (age, weight, sex, race, baseline CP, and CH50) were statistically evaluated using stepwise forward selection / backward elimination. During stepwise forward selection, covariates that contributed at least 6.63 units of change in the minimum value of the objective function (MVOF) (α = 0.01, one degree of freedom) were considered statistically significant. The covariates that contributed the most significant change in the MVOF (minimum p-value) were included in the new base covariate model. The new models were also evaluated for reduction in inter-individual variability (IIV) of the PK parameters. Highly correlated covariates were not included in the single covariate-parameter submodel. This process was repeated until no further covariates produced a significant change in MVOF. The resulting model was considered a full multivariable model. The appropriateness of the structural and statistical models was fully evaluated and refined as needed.

[0296] In backward elimination of covariates, each covariate was removed from the parameter equation individually. A covariate was considered significant if it contributed at least 10.83 changes in the MVOF value (α = 0.001, one degree of freedom) when removed from the model. The least significant covariate (highest p value > 0.001) was removed from the model, and this reduced model was then used as the new base multivariate model. The backward elimination procedure was repeated until all remaining covariates were statistically significant (p < 0.001).

[0297] Statistical analysis

[0298] Safety: Safety analyses (including AEs, laboratory parameters, vital signs, and ECGs) were based on review of individual values ​​and descriptive statistics. The treatment-emergent adverse event (TEAE) time period was defined as the time from the first SAR445088 administration to the end-of-study visit (inclusive). AEs were coded according to the Medical Dictionary for Regulatory Activities, version 23.0, and severity was graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events, version 5.0. The number (%) of participants experiencing TEAEs was summarized by dose level group. Potentially clinically significant abnormalities in clinical laboratory test results, vital signs, and ECGs were labeled and summarized by dose level group using frequency tables.

[0299] The SLE panel test was evaluated by summarizing the number (%) of participants with negative, positive or marginal status data for each dose level group, parameter and visit. The number of participants who experienced local injection site reactions (e.g., pain, tenderness, erythema, swelling / nodules or itching) was summarized by dose level group and grade (mild / moderate, severe and very severe). The immunogenicity of SAR445088 was evaluated by summarizing the number and percentage of participants with ADA positive or negative status by dose level group and visit. In the case of positive ADA, descriptive statistics were used to summarize absolute ADA concentrations by dose level and visit.

[0300] Pharmacokinetics: Descriptive statistics were used to summarize SAR445088 PK parameters by dose level. The maximum drug concentration in plasma (C max ), the area under the curve (AUC last ) or the area under the curve (AUC 0-168 Dose proportionality was assessed using a power model of AUC for each route (IV and SC). Accumulation was assessed using a linear model of log-transformed cumulative ratios. The log-transformed C max and AUC 0-168 The variance component of .

[0301] Pharmacodynamics: Percent CP activity, percent AP activity, CH50, and C4 were analyzed in all participants. Descriptive statistics were provided by dose level and measurement time.

[0302] result

[0303] Demographics and disposition of participants

[0304] Overall, 93 healthy participants were recruited in Parts 1 and 2. In Part 1, a total of 65 participants were recruited. A single dose of SAR445088 or matching placebo IV was administered to a total of 45 participants, including 33 participants who received SAR445088 and 12 participants who received placebo. A single dose of SAR445088 or matching placebo SC was administered to a total of 20 participants, including 15 participants who received SAR445088 and 5 participants who received placebo. In the Japanese cohort, three participants received SC doses of SAR445088. All cohorts completed recruitment of eight participants (six SAR445088 and two placebo) as planned, except for the 50mg / kg IV non-Japanese cohort and the 300mg SC Japanese cohort. Recruitment in these two cohorts was terminated early due to recruitment difficulties during the COVID-19 pandemic. One patient in the 300mg SC Japanese discontinued the study due to non-study related reasons.

[0305] In Part 2, a total of 28 participants were recruited and randomized to receive multiple doses of SAR445088. Twenty-two participants received SAR445088 and six participants received placebo. The subject taking placebo withdrew from the study and discontinued study treatment due to a TEAE (decreased neutrophil count); this subject was not replaced. Recruitment in the 600 mg SC once daily × 3 days cohort was terminated early due to recruitment difficulties during the COVID-19 pandemic. All recruited participants in Parts 1 and 2 were included in the safety, PK, and PD analyses. A flowchart of the disposition of patients is shown in Figure 1 The demographic and baseline characteristics are provided in Tables 2-4.

[0306] Table 2: Summary of demographic characteristics of the study population (Part 1, IV)

[0307]

[0308]

[0309] Table 3: Summary of demographic characteristics of the study population (Part 1, SC)

[0310]

[0311]

[0312] Table 4: Summary of demographic characteristics of the study population (Part 2)

[0313]

[0314]

[0315] Safety and Tolerability

[0316] 93 randomized healthy adult participants who received at least one dose of SAR445088 or matching placebo were included in the safety population. No deaths, severe TEAEs, or treatment-emergent serious adverse events (TESAEs) were reported during the study.

[0317] Overall, no relevant trends were observed in vital signs, ECG, or laboratory values. No serious infections or meningococcal infections occurred.

[0318] Pharmacokinetics

[0319] The mean (±SD) PK parameters following IV and SC dosing and the corresponding plasma concentration-time profiles for Parts 1 and 2 are presented in Table 5 and Figure 2, respectively.

[0320] Table 5: Summary of mean (±SD) PK parameters

[0321]

[0322]

[0323] Decline; t max , the time to reach the maximum drug concentration; V d F, apparent volume of distribution

[0324] a For the IV cohort, the median infusion time was 0.6 h, and the first post-dose sample was collected 1 h after the end of the infusion.

[0325] b Expressed as median (range).

[0326] c The extrapolation ratios ranged from 28.3% to 48.9%, and the estimated parameters (AUC, CL, CL / F, V z and V z / F) should be interpreted with caution.

[0327] d Apparent clearance (CL / F) after subcutaneous doses was estimated.

[0328] e Estimation of the apparent volume of distribution after a subcutaneous dose (V d / F).

[0329] f Estimated t after last dose max : Median (minimum, maximum).

[0330] g The cumulative ratio of weekly dosing was calculated as the AUC on day 22. 0-168h / Day 1 AUC 0-168h ratio.

[0331] The bioavailability of SAR445088 after SC dosing ranged from 67.5% to 92.0%. After a single IV or SC dose of SAR445088, exposure increased in a dose-proportional manner across the range of 2 to 50 mg / kg IV or 600 to 1200 mg SC. After a single SC dose, the PK profile of SAR445088 was described by slow absorption (3.95–21.1 days). A long half-life after IV or SC administration was observed (cohort mean: 8–15 weeks).

[0332] Treatment-related ADA were observed in two participants in Part 1 (one subject each after 2 mg / kg IV and 600 mg SC) and in two participants in Part 2 (300 mg SC group), with no apparent effect on SAR445088 exposure.

[0333] Pharmacodynamics

[0334] Percent CP activity: Dose-dependent inhibition of percent CP activity was observed after single or multiple doses of SAR445088, analyzed as change relative to pre-dose levels ( Figures 3A-3C ). In Part 1, the maximum inhibition of percent CP activity relative to baseline was >90% on Day 1 after a single 30 or 50 mg / kg IV dose, and the maximum inhibition of percent CP activity relative to baseline was 55% on Day 7 after a single 1200 mg SC dose. The extent and duration of inhibition of percent CP activity after a single IV or SC dose appeared to be dose-dependent. The maximum inhibition of percent CP activity of >90% persisted for approximately 4 weeks after 50 mg / kg IV administration. In Part 2, the maximum inhibition of percent CP activity relative to baseline was 82% on Day 35 after the fourth weekly 600 mg SC dose, and the inhibition of percent CP activity persisted for the duration of the study period. After the third daily 600 mg SC dose, the maximum inhibition of percent CP activity relative to baseline was 70.5% on Day 15 and declined thereafter.

[0335] CH50 inhibition: Dose-dependent inhibition of CH50 was observed after single or multiple doses of SAR445088 ( Figure 3D-3F ). In Part 1, a sustained decrease in CH50 (≤10 IU / mL) was observed within 1 h with the 16 mg / kg IV dose and persisted at this level until Week 3. A longer sustained decrease in CH50 to <10 IU / mL was observed with administration of 30 mg / kg IV (>3 weeks) or 50 mg / kg IV (>8 weeks). Following a single 1200 mg SC dose, a decrease in CH50 (≤10 U / mL) was observed on Day 5, which persisted until Day 15. In Part 2, a decrease in CH50 (≤10 U / mL) was observed in the 600 mg SC weekly cohort on Day 22, which persisted at this level until Day 84. Daily administration of 600 mg SC for 3 days resulted in a decrease in CH50 to ≤10 IU / mL on Day 8, which persisted until Day 22. There was no change in percent AP activity following a single IV or SC dose and was similar to baseline levels throughout the study period for all participants. C4 is the first complement component cleaved by activated C1s in CP. Total C4 levels did not change after single or multiple doses and were similar to baseline levels throughout the study period in all study participants. Without wishing to be bound by theory, this may be due to the fact that this study was conducted in healthy participants with only low levels of C1s activation.

[0336] Population PK / PD Modeling

[0337] Population PK Modeling: A total of 70 participants (who received SAR445088) with 1244 concentration records were used for model development. The population PK of SAR445088 was characterized by a two-compartment model with first-order absorption and linear elimination. The typical clearance was 2.12 mL / h. The total volume of distribution (V d )(5.45L) is consistent with plasma volume, and drug distribution is confined to the blood circulation. The model-estimated bioavailability after SC administration was 75.4%. Population PK parameters were estimated with good accuracy, with relative standard errors (RSE) ≤ 30%. The magnitude of the estimated IIV was moderate (36.6% CV to 75.0% CV). Results of the visual predictive check (VPC) showed that most observed concentrations were within the predicted range (5th to 95th percentiles). Among the covariates tested, the effect of body weight on peripheral chamber volume (V2) was retained as a statistically significant covariate. As body weight increased from 50.4kg (minimum) to 100.8kg (maximum), peripheral V d Increased by 2.3 times ( Figure 4A ).

[0338] Exposure-Response Relationship: A total of 93 participants (receiving SAR443088 or placebo) with 1496 CP and 963 CH50 records were used to develop the PK / PD model. The base CP PK / PD model was a direct response model with nonlinear sources of variability. Baseline CP activity was identified as the major source of SAR445088 CP population pharmacokinetic-pharmacodynamic (PopPK / PD) variability. The observed CP baseline ranged from 66.2% to 186%, while the modeled base CP ranged from 75.3% to 139%. VPC showed that most observed concentrations were within the predicted range (5th to 95th percentiles). For the CP percentages, typical IC 50 is 96.4 μg / mL, and the typical IC 90 The basic CH50 PK / PD model is a direct response model with nonlinear PD effects ( Figure 4B-4E ). No covariates were identified for the CH50 population PK / PD model that met the pre-specified inclusion criteria. The results of the VPC showed that most observed concentrations were within the predicted range (5th to 95th percentiles). For CH50, the typical IC 50 is 16.6 μg / mL, and the typical IC 90 It is 57.0μg / mL.

[0339] Using the final CP model, the concentration-time profile of SAR445088 was simulated for a patient population (n=200) weighing 70 kg for the proposed treatment regimen (50 mg / kg IV loading and 600 mg SC weekly maintenance). The proposed treatment regimen achieved inhibition of CP after the first dose (based on Wieslab CP determination PK / PD relationship) and maintained this inhibition throughout the treatment period.

[0340] discuss

[0341] In this FIH trial in healthy participants, SAR445088 was well tolerated at single or multiple doses up to 50 mg / kg IV, and no safety issues were observed. All reported TEAEs were mild or moderate in severity. No deaths, serious AEs, serious infections, encapsulated bacterial infections, or meningitis were reported in this study. Because SAR445088 keeps the alternative and lectin pathways of complement intact, the risk of encapsulated bacterial infections due to administration of SAR445088 may be lower than other complement inhibitors. Vaccination against encapsulated bacteria (such as meningococci) is required in this study to mitigate this risk. Based on data from participants with congenital CP defects, a theoretical risk of C1s inhibition is the development of SLE. In this study, no AEs related to the development of SLE were reported.

[0342] The PK of SAR445088 in healthy participants is characterized by a long elimination half-life after IV or SC administration, which is partly due to the methionine to leucine mutation (M428L) and asparagine to serine mutation (N434S) increasing the neonatal Fc receptor (FcRn) binding affinity at pH 6.0. In addition, SAR445088 only binds to activated C1s. Target-mediated drug disposal. Target-mediated drug disposal (TMDD) was not observed at the dose level studied. Good bioavailability after SC administration was also observed, which provides the potential for self-administration at home for chronic diseases that require maintenance treatment. An approximate dose-proportional increase in SAR445088 exposure was observed within the dose range evaluated.

[0343] The incidence of treatment-induced ADA was low and had no apparent impact on SAR445088 exposure, PK parameters, or the safety profile of SAR445088.

[0344] Complement inhibition was measured using two different assays (Wieslab CP and CH50 hemolytic assay). For each assay, a dose-dependent inhibition of CP was observed that reached levels observed in individuals with congenital or acquired classical complement deficiency. In addition, when comparing the Wieslab CP and CH50 hemolytic assays, an IC 50 Given the approximately 8-fold difference in values, a higher drug exposure was required to achieve the same level of CP inhibition with the Wieslab CP assay compared to the CH50 hemolytic assay. This may be due to differences in sample assay dilution and readout in the CH50 and Wieslab CP assays. Of note, in vitro studies using SAR445088 in patient blood samples also yielded similar differences between CP and CH50 percentages.

[0345] in conclusion

[0346] In this FIH trial in healthy adults, SAR445088 was well tolerated and no serious or severe adverse events occurred. In healthy participants, SAR445088 demonstrated dose-dependent inhibition of classical CP after a single IV or SC and multiple weekly or daily SC ascending doses.

[0347] Example 2. Multiplex Evaluation of the Safety, Tolerability, and Activity of SAR445088 in Adults with Cold Agglutinin Disease Center, Phase 1b, open-label, non-randomized, single-dose study

[0348] Rationale

[0349] The purpose of this Phase 1b study is to determine the safety, tolerability, activity, and PK of a single IV dose of SAR445088 in an adult patient population with cold agglutinin disease (CAD). By testing SAR445088 in CAD, this study aims to detect biomarker evidence of improved hemolysis to support proof of concept for SAR445088.

[0350] Methodology

[0351] This was a multicenter, Phase 1b, open-label, non-randomized study evaluating the safety and tolerability of a single IV dose of SAR445088 in adult participants with cold agglutinin disease (CAD).

[0352] This study is designed to evaluate the effect of SAR445088 on classical complement pathway-mediated hemolysis in CAD and to generate data to determine dosing for future studies. The study design includes up to three single-dose IV cohorts. Dosing is initiated in Cohort 1a at a dose of 30 mg / kg IV, and study decisions regarding dose selection for the next cohort and / or within-cohort expansion are made based on clinical response (total bilirubin and hemoglobin) and variability in response between participants. Following Cohort 1a, either a higher-dose cohort (Cohort 1b) or a lower-dose cohort (Cohort 1c) may be recruited. Cohort 1b was not enrolled, while Cohort 1c was enrolled at a dose of 15 mg / kg IV.

[0353] Number of participants

[0354] Planned: In any IV cohort, approximately 6 participants will be dosed.

[0355] Treated: Twelve adult participants were treated with a single IV dose of SAR445088. Six participants received 30 mg / kg IV, and 6 participants received 15 mg / kg IV.

[0356] All 12 treated participants were included in all analysis populations (safety, pharmacodynamics, pharmacokinetics, ADA, BA, PK / PD, and PK / BA)

[0357] The main criteria for diagnosis and inclusion and exclusion are:

[0358] Male and female participants were aged 18 years and older with CAD and a hemoglobin level ≤11 mg / dL at screening.

[0359] i) Has been previously vaccinated against enveloped organisms

[0360] ii) No systemic corticosteroids (more than 10 mg / day of prednisone or its equivalent), systemic immunosuppressants, systemic cytotoxic agents, specific complement system inhibitors other than SAR445088, or anti-CD20 therapy within three months before screening

[0361] iii) Not received any biological agents (antibodies or their derivatives) within 4 months before screening.

[0362] Duration of study intervention

[0363] The total study duration for each participant was approximately 23 weeks:

[0364] Screening: Maximum 56 days, minimum 14 days (Day -56 to Day -1).

[0365] Treatment: 1 day (Day 1); and,

[0366] • Follow-up and end of study: 105 days after SAR445088 administration (follow-up from Day 2 to Day 71; end of study visit on Day 106).

[0367] The study was considered complete for each participant when the participant completed all scheduled study procedures.

[0368] Statistical methods

[0369] Security:

[0370] Safety analyses (AEs, laboratory parameters, vital signs, ECG) were based on review of individual values, and descriptive statistics, and focused on the TEAE period, defined as the time from first IMP administration to end of study visit (inclusive). All safety analyses were performed using the safety population by dose level cohort.

[0371] Adverse events were coded according to the Medical Dictionary for Regulatory Activities (MedDRA Version 24.1). Their severity was graded according to NCI-CTCAEv5.0. The number (%) of participants experiencing TEAEs was summarized by dose level cohort. Potentially clinically significant abnormalities (PCSA) in clinical laboratory test results, vital signs, and ECG were marked and summarized by dose level cohort using a frequency table. The SLE panel test was evaluated by summarizing the number (%) of participants with negative, positive, or marginal status data for each dose level cohort, parameter, and visit. Quantitative SLE parameters were summarized using descriptive statistics (N, mean, SD, SEM, median, minimum, and maximum) and provided by dose level cohort and measurement time. The number of participants experiencing local injection site reactions (pain, tenderness, erythema, swelling / nodules, or pruritus) was summarized by dose level cohort and grade (mild / moderate, severe, and very severe).

[0372] Anti-SAR445088 Antibody (ADA):

[0373] Anti-SAR445088 antibodies (anti-drug antibodies, ADA): The immunogenicity of SAR445088 was assessed using the ADA population by evaluating the ADA-positive or -negative status of the participants and summarized by dose level cohort and visit. In addition, if ADA status was confirmed, the absolute ADA concentrations were summarized in descriptive statistics by dose level cohort and visit.

[0374] Pharmacokinetics (PK):

[0375] Descriptive statistics were used to summarize the calculated SAR445088 concentrations in plasma by dose level cohort. In addition, summary plots of the raw data (mean + / - SD) were provided, with one curve for each dose level cohort. The population PK model developed in healthy subjects was evaluated in CAD patients by statistically evaluating the model-predicted SAR445088 concentrations for each participant versus the observed SAR445088 concentrations.

[0376] Pharmacodynamics (PD):

[0377] All PD analyses were performed using the PD population by dose level cohort. For all participants included in the PD population, the quantitative parameters percentage CP activity, percentage AP activity, CH50, and total C4 were analyzed as raw data, changes from baseline, and percentages normalized to baseline. The percentage CP inhibition was analyzed as raw data. Descriptive statistics on the raw data, changes from baseline, and percentages normalized to baseline for percentage CP activity, CH50, and total C4 were provided by dose level cohort and measurement time. For all PD parameters (percent CP activity, percentage CP inhibition, CH50, and total C4), time curves (mean ± standard error of the mean [SEM]) and changes from baseline for each dose level cohort are provided.

[0378] Biomarker Activity (BA)

[0379] Hematologic BAs for all participants included in the BA population were analyzed as raw data and changes from baseline. For all BA parameters and changes from baseline, time profiles for each dose level cohort are provided (mean ± standard deviation [SD]).

[0380] PK / PD and PK / BA

[0381] The relationship between PD and BA variables and SAR445088 plasma concentrations was explored graphically using the PK / PD population and the PK / BA population. The mean (± SEM) changes of PD and BA data relative to baseline and the mean SAR445088 plasma concentration versus time (hours after dosing) were superimposed on the same graph. Scatter plots of PK SAR445088 concentrations versus PD raw data and changes relative to baseline were provided over time.

[0382] Summary of Results and Conclusions:

[0383] A total of 12 participants received a single dose of SAR445088 via IV administration.

[0384] Demographics and other baseline characteristics:

[0385] The age range of the study participants was 54 to 80 years (mean, 67.4; median, 69.0). This study included 1 male participant and 11 female participants.

[0386] Exposure:

[0387] The study started with 3 participants receiving a single dose of 30 mg / kg IV in Cohort 1a. After dosing, 2 of the 3 participants were found to have bilirubin within the normal range at baseline, even though their bilirubin was above ULN during screening. It was therefore decided to enroll an additional 3 participants at the 30 mg / kg dose. In Cohort 1a, it was observed that the bilirubin levels of the 4 participants with baseline bilirubin levels above ULN normalized by Day 8. It was therefore decided to proceed to Cohort 1c at the 15 mg / kg dose.

[0388] Overall, 6 participants received a single dose of SAR445088 30 mg / kg IV and 6 participants received a single dose of SAR445088 15 mg / kg IV.

[0389] Safety Results:

[0390] There were no treatment-emergent SAEs, TEAEs leading to death, or TEAEs leading to permanent study discontinuation. No participant experienced serious infections, meningococcal infections, allergic and / or hypersensitivity reactions, or thromboembolic events.

[0391] Overall, SAR445088 was generally well tolerated. No new safety issues were identified.

[0392] Pharmacodynamic results:

[0393] After a single dose of SAR445088 30 mg / kg IV, the mean percentage CP activity decreased from 49.70% at baseline to <10% at D1 1H, followed by a gradual increase in CP activity. The mean CH50 value decreased from 21.7 IU / mL at baseline to <10 IU / mL at D11H. At the EOS visit, the mean CH50 value remained <10 IU / mL. After a single dose of SAR445088 15 mg / kg IV, the mean percentage CP activity decreased from 56.09% at baseline to 10.23% at D1 1H, followed by a gradual increase in CP activity. The mean CH50 value decreased from 25.7 IU / mL at baseline to <10 IU / mL at D1 1H. At the EOS visit, the mean CH50 value was 13.6 IU / mL. No decrease in AP activity was observed over the duration of the study.

[0394] The effects of SAR445088 on clinical biomarkers of hemolysis were evaluated by time point using mean total bilirubin (a marker of red blood cell destruction) and mean hemoglobin. After a single dose of 30 mg / kg, mean bilirubin decreased from 21.18 μmol / L at baseline (upper limit of normal 17.1 μmol / L) to a trough of 7.35 μmol / L at D29 and 9.41 μmol / L at the EOS visit. After a single dose of 15 mg / kg, mean bilirubin decreased from 35.91 μmol / L at baseline to a trough of 11.97 μmol / L at D71 and a value of 20.52 μmol / L at the EOS visit ( Figure 5 ).

[0395] After a single dose of 30 mg / kg, hemoglobin increased from a mean of 97.3 g / L at baseline to a peak of 121.7 g / L at D43, and the value at the EOS visit was 121.3. After a single dose of 15 mg / kg, hemoglobin increased from a mean of 85.9 g / L at baseline to a peak of 121.5 g / L at D57, and the value at the EOS visit was 119.5 g / L ( Figure 6 ). In the 15 mg / kg group, two participants received transfusions of packed red blood cells on the day of SAR445088 administration. The analysis of hemoglobin increases from baseline in this group may have been affected by these transfusion events.

[0396] Pharmacokinetic results:

[0397] Based on the PopPK model developed in healthy participants, SAR445088 concentrations were generally within the 90% prediction interval, suggesting that the PK in CAD participants was similar to that in healthy participants ( Figure 7 ).

[0398] Other results:

[0399] No participants had treatment-induced ADA, treatment-enhanced ADA, or treatment-emergent ADA. No participants had indeterminate ADA.

[0400] in conclusion:

[0401] A single dose of SAR445088 had an acceptable safety and tolerability profile in adult participants with CAD. During the 15-week follow-up period, a sustained decrease in CH50 was observed after a single dose of 30 mg / kg IV and 15 mg / kg IV. The observed decrease in CH50 values ​​corresponded to a decrease in total bilirubin and an increase in hemoglobin during the 15-week follow-up period. Overall, these findings suggest that a single IV dose of 30 or 15 mg / kg SAR445088 results in classical complement inhibition, hemolysis control, and anemia improvement in adult patients with CAD.

[0402] Example 3. SAR445088 Dose Regimen Selection for Phase 2 / 3 Study

[0403] The primary objective of this study was to mimic the dosing regimen of a Phase 2 / 3 study in patients with cold agglutinin disease (CAD) and chronic inflammatory demyelinating polyneuropathy (CIDP).

[0404] Methodology:

[0405] software

[0406] SAR445088 concentration versus time data were simulated using NONMEM software version 7.4.1 (ICON, Development Solutions, Elliot City, MD, USA).

[0407] Simulated data sets, graphical analyses, descriptive statistics, and evaluation of NONMEM output were performed using R (R Core Team. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing. Vienna, Austria, 2019).

[0408] Population pharmacokinetic / pharmacodynamic models

[0409] PopPK and PopPK / PD models in healthy subjects

[0410] Using data from single and multiple ascending dose studies in healthy adult participants, a population pharmacokinetic (PopPK) model and a population pharmacokinetic / dynamic (PopPK / PD) model (Example 1) of SAR445088 were developed. These studies investigated 2 to 50 mg / kg intravenous (IV) doses and 300 to 600 mg subcutaneous (SC) doses as single or repeated doses.

[0411] The pharmacokinetics (PK) of SAR445088 in healthy subjects was well described by a linear elimination two-compartment model with first-order absorption after subcutaneous (SC) administration. The typical clearance (CL) was 2.21 mL / h. The total distribution volume (V) was consistent with the plasma volume, indicating that the drug distribution was confined to the blood circulation. The peripheral distribution volume (V p ) is the main source of PK variability for SAR445088. As body weight increases from 50.4 kg to 100.8 kg (the range observed in studies in healthy subjects), V p In addition, all other covariates, including age, sex, race, and anti-drug antibody (ADA) status, were evaluated and none was found to have a statistically significant effect on SAR445088 PK.

[0412] The popPK model showed that SAR445088 was distributed more in organs and tissues in individuals with higher body weight. The rapid intercompartmental clearance (Q = 14 mL / h) and slow CL indicated that SAR445088 spent most of its time in the central volume of distribution (V c Therefore, it is expected that body weight will have a negligible effect on SAR445088 plasma concentrations within the tested range.

[0413] PopPK / PD models were developed for complement pathway activity as measured by the Wieslab classical pathway (CP) and 50% hemolytic fraction (CH50) as pharmacodynamic (PD) markers. Differences were observed in the PK / PD of SAR445088 on CP and CH50: IC of CP 90 The IC value of CH50 is 458 μg / mL. 90 As a conservative approach, the CP PK / PD model was used to propose dosing regimens for Phase 2 CAD and CIDP studies.

[0414] Simulations for dose selection for Phase 2 / 3 studies

[0415] PopPK and popPK / PD models were used to simulate different dosing schedules (once a week, qw, every other week, q2w, monthly, qm, every 12 weeks, q12w) with or without an IV or SC loading dose, with the goal of achieving a CP less than 10% (i.e., >90% inhibition) or a CH50 <10 U / mL.

[0416] Dosing Regimen Selection for Phase 2 CAD and CIDP Studies

[0417] Based on the popPK / PD model for CP described above, single IV doses of 15 and 30 mg / kg were selected for a proof-of-concept study in CAD patients (Example 2). These models were also used to select a loading dose of 50 mg / kg IV, followed by 600 mg SC weekly starting on day 8 for a Phase 2 CIDP study.

[0418] Constant-dose versus weight-based regimens

[0419] The popPK model for SAR445088 was used to simulate PK by body weight. As a conservative approach to evaluate the effect of body weight on SAR445088 exposure, the PK parameters CL and V were allometrically scaled based on body weight. The exponent used for CL was 0.85, and the exponent used for V was 1. The equations for CL and V are as follows:

[0420]

[0421] According to Table 6, the body weight distribution from 35 to 200 kg was classified into zones for easy comparison.

[0422] Table 6. Weight divisions

[0423]

[0424] From the weight distribution, patients (n=1000) were randomly selected with replacement using an R script. For each patient (n=1000), the SAR445088 exposure profile was simulated to steady state. The PK parameter area under the curve (AUC) at steady state was estimated. ss ) and the pre-dose concentration at steady state (C trough ).

[0425] Weight distribution and simulation

[0426] The PK / PD model of CH50 was used to evaluate the following regimens matched to 70 kg individuals in the CAD and CIDP patient populations:

[0427] Constant dosing regimen: 3.5 g IV q12w, with an additional 3.5 g IV dose on day 29

[0428] Weight-based dosing regimen: 50 mg / kg q12w, with an additional 50 mg / kg IV dose on day 29.

[0429] Weight distribution data were obtained from the following studies:

[0430] 1) CAD: Cadenza (BIVV009-03) and Cardenal (BIVV009-04) studies; n=66, median=65.6+ / -13.5 kg.

[0431] 2) CIDP: CIDP patient population from the Centers for Disease Control database (Centers for Disease Control and Prevention. US CIDP patient weight. February 2022); n=100, median=83.9+ / -35.8 kg.

[0432] The population density of the indications overlapped, with the majority of patients being in the 50 to 90 kg range.

[0433] Results and discussion:

[0434] Dose Regimen Selection for Phase 2 CAD and CIDP Studies

[0435] CAD

[0436] Based on the PK / PD model of CP, a single IV dose of 30 mg / kg was predicted to achieve a maximum concentration of ∼660 μg / mL in CAD patients (C max ), CP was less than 10% in 60% of patients ( Figure 8 ).

[0437] CIDP

[0438] Loading dose of 50 mg / kg IV followed by 600 mg SC qw on day 8 Predicted steady-state predose concentration (C trough ) was approximately 1210 μg / mL, and during the entire treatment period, most patients (50%) had CP values ​​below 10% ( Fig. 9 ). Based on AUC ss Comparative 26-week non-human primate (NHP) studies with no observed adverse effect (NOAEL) (AUC0-12week = 8,388,000 μg*h / mL) had an estimated exposure margin greater than 2.

[0439] A loading dose is necessary to rapidly achieve 90% CP inhibition. For example, a 1200 mg SC dose administered every 2 weeks (q2w) is sufficient to achieve 90% CP inhibition in patients with 1145 μg / mL C trough 90% CP suppression was achieved at approximately 12 weeks of treatment.

[0440] Alternatives

[0441] Lower dose

[0442] The dose proposed for phase 2 CIDP is based on CP PK / PD as a conservative approach. For SC and IV, a regimen based on CH50 PK / PD is proposed, which requires a lower maintenance dose.

[0443] SC. The proposed lower maintenance dose regimen for SC administration is 50 mg / kg IV on day 1 and 300 mg SC qw on day 8. Based on the CH50 popPK / PD model, this regimen achieves rapid SAR445088 levels that reduce CH50 values ​​to below 10 U / mL and maintains a C of ~600 ug / mL for the duration of treatment. trough Continue to exist.

[0444] IV. The proposed lower maintenance dose regimen for IV administration is 50 mg / kg IV q8w on day 1. Based on the CH50popPK / PD model, this regimen achieves rapid SAR445088 levels that reduce CH50 to below 10 U / mL and maintains a C of ∼800 g / mL for the duration of treatment. trough Continue to exist.

[0445] Constant-dose versus weight-based regimens

[0446] A weight-based regimen (50 mg / kg IV dose every twelve weeks (q12w) with an additional 50 mg / kg IV dose on day 29) was compared to an equivalent constant dose in 70 kg individuals to support dose selection for the CAD Phase 3 study. Fig.10 A simulation of 3.5 g IV every twelve weeks with an additional 3.5 g IV dose on day 29 is shown in FIG. At steady state, 12% of patients are predicted to have SAR445088 levels below 300 μg / mL, but 100% of patients are predicted to have SAR445088 levels above 100 μg / mL. For the proposed regimen, the C between 3.5 g and 50 mg / kg trough The comparison shows that C trough The two methods were comparable (Table 7).

[0447] Table 7. PopPK simulations of constant dose regimens versus weight-based dose regimens C trough : 3.5 g or 50 mg / kg IV q12w starting on day 1 + 3.5 g or 50 mg / kg IV on day 29

[0448]

[0449] The simulated area under the curve (AUC) at steady state ss ) is presented in Fig.11 The AUC values ​​were calculated and summarized in Table 8. For the constant dose method, AUC ss There was a trend toward a decrease from low body weight (partition A) to high body weight (partition H); this decrease was ~2.8-fold. For the weight-based dosing regimen, AUC ss There was a trend of increasing weight from low weight (partition A) to high weight (partition H); this increase was -1.2-fold.

[0450] Table 8. SAR445088 Constant Versus Weight-Based Dose Simulated Steady-State AUC

[0451]

[0452] For the middle weight partition of 65-105 kg, exposures between the 3.5 g constant and 50 mg / kg dosing regimens were comparable and were expected for most patients. The greatest differences were observed for very low (Partition A) and very high (Partition H) weights, where constant and weight-based AUC ss The ratio is 1.54 for partition A and 0.46 for partition H.

[0453] Simulated minimum plasma drug concentration at steady state (C trough ) presented in Fig.12 and summarized in Table 9. For the 3.5 g constant dose method, C trough There was a trend toward a decrease from low body weight (partition A) to high body weight (partition H); this decrease was ~2.3-fold. For the 50 mg / kg dosing regimen, C trough There was a trend toward an increase from low body weight (partition A) to high body weight (partition H); this increase was ~1.5-fold. ss Similarly, the intermediate partitions (C to F) were comparable between the constant dose approach and the weight-based dose approach. The greatest differences were observed for very low (Partition A) and very high (Partition H) weights, with the constant and weight-based C trough The ratio is 1.56 for partition A and 0.46 for partition H.

[0454] Table 9 - SAR445088 constant dose vs. body weight based dose simulated steady state C trough

[0455]

[0456] The steady-state exposure (AUC ss ) were compared with the NOAEL established in the 26-week NHP toxicity study (Table 10). All weight partitions had an exposure margin greater than 3.

[0457] In the healthy subject study (Example 1), the highest overall exposure was from the 50 mg / kg IV single dose cohort: C max is 1180 μg / mL, and AUC 0-∞ The proposed constant dose regimen exposure would be 1.73–0.62 times the exposure observed in healthy subjects at 35-200 kg.

[0458] Table 10. Exposure Margins for Constant Dose Regimens vs. Weight-Based Dose Regimens vs. NHP NOAELs

[0459]

[0460] For a constant dose, C trough The analysis of the levels is presented in Table 11, and for the weight-based dosing approach, C trough The analysis of the levels is presented in Table 12.

[0461] Table 11. SAR445088 constant dose simulation C trough analyze.

[0462]

[0463] Table 12. Dose simulation C of SAR445088 based on body weight trough analyze

[0464]

[0465] At steady state, 92% of patients are predicted to have a C above 300 μg / mL. trough For the constant-dose approach, 84 of 982 patients were below 300 μg / mL and 18 of 982 patients were below 200 μg / mL; this was comparable to the weight-based dosing approach, where 82 of 982 patients were below 300 μg / mL and 12 of 982 patients were below 200 μg / mL. Overall, 100% of patients were predicted to be above 100 μg / mL (above IC 90 2 times).

[0466] in conclusion:

[0467] 1) Bayesian analysis showed that CAD patients had a PK profile similar to that of healthy participants.

[0468] 2) The decrease in CH50 values ​​after SAR445088 exposure in CAD patients was associated with improved clinical markers bilirubin and hemoglobin, confirming that CH50 is a relevant PD marker for SAR445088.

[0469] 3) The dose regimens selected for the Phase 2 study are predicted to achieve CP less than 10% (i.e., >90% inhibition) or CH50 <10 IU / mL in most patients throughout the treatment period. The predicted exposure margins for these regimens are greater than 2 compared to the 26-week NHP NOAEL.

[0470] 4) The constant dose regimen appears to produce C values ​​comparable to the weight-based regimen in the body weight range of 35 to 200 kg. trough C with similar values trough The predicted exposure margin for the 35 to 200 kg constant dose regimen compared to the 26-week NHP NOAEL was greater than 3.

[0471] Example 4. A Phase 2, Multicenter Trial to Evaluate the Efficacy, Safety, and Tolerability of SAR445088 in Adults with CIDP Open-label, non-randomized, proof-of-concept study

[0472] As described in Example 1 above, a single SAR445088 IV dose of 30 mg / kg or 50 mg / kg resulted in a maximum CP inhibition of at least 90%. The 90% cutoff for CP inhibition was derived from a previous study of first generation C1s mAbs for cold agglutinin disease (CAD) that found 90% CP inhibition to be a predictor of therapeutic efficacy (blocking hemolytic activity in the blood compartment) ( et al. Blood. [Hematology] 2019 Feb 28;133(9):893-901. doi:10.1182 / blood-2018-06-856930. Epub 2018 Dec 17). Therefore, this degree of CP inhibition is expected to translate into a therapeutic benefit (improvement of severe hemolytic anemia in CAD). CAD represents a good model for complement-mediated diseases in the blood compartment. However, the target site in CIDP is in the peripheral nerves, which requires SAR445088 to cross the blood-nerve barrier. To account for uncertainty in the ability of SAR445088 to achieve adequate exposure at this target site in CIDP patients, twice the CP IC90 plasma concentration (i.e., 1200 μg / mL) was used as the exposure criterion for Phase 2 dose selection.

[0473] Available data from the Phase 1 study support the initiation of a Phase 2 study in patients with CIDP, using a dose expected to result in at least 90% inhibition of the CP target, which is expected to reverse pathological activation of this pathway. This effect could induce clinical benefits associated with reversal of neuroinflammation, remyelination, and prevention of axonal degeneration.

[0474] The purpose of this SAR445088 Phase 2 study is to determine the preliminary efficacy, safety, and tolerability of SAR445088 in three CIDP subpopulations, including those treated with standard of care (SOC), those refractory to SOC, and those naive to SOC. Results from this Phase 2 study determine proof of concept (PoC) and inform the Phase 3 trial.

[0475] This study is a global, multicenter, Phase 2, open-label, PoC study that evaluates the efficacy, safety, and tolerability of SAR445088 administered to 3 CIDP patient subpopulations: patients being treated with SOC therapy defined as intravenous immunoglobulin (IVIg), subcutaneous immunoglobulin (SCIg), or corticosteroids; patients with CIDP refractory to SOC; and patients with CIDP who are not receiving SOC treatment. This study will consist of two parts: an initial 24-week treatment period (Part A), followed by an optional extension period (Part B) designed to evaluate long-term safety and tolerability, which will provide up to 1 year of additional treatment. A schematic diagram of the study is shown in Fig.13 middle.

[0476] This study enrolled patients from three CIDP subgroups separated according to their experience with SOC therapy (defined as immunoglobulin or corticosteroids):

[0477] I. Refractory to SOC (SOC refractory); n = 20 participants: patients with evidence of SOC failure or inadequate response (defined as no clinically meaningful improvement and persistent INCAT score ≥ 2 after a minimum of 12 weeks of SOC treatment prior to screening), or patients who are unable to receive or continue immunoglobulin or corticosteroid treatment due to side effects. Patients should not receive immunoglobulin (IVIg or SCIg) within 12 weeks prior to screening. Patients have INCAT scores: 2-9 (score 2 should be entirely from the leg disability component of INCAT). Clinically meaningful improvement (and deterioration) is defined as one of the following: a decrease (increase) of ≥ 1 point in the adjusted INCAT score, an increase (decrease) of ≥ 4 points in the I-RODS total score, an increase (decrease) of ≥ 3 points in the MRC total score, an improvement (decrease) of ≥ 8 kPa in the mean grip strength (one hand), or an equivalent improvement (deterioration) based on information documented in the medical record and according to the judgment of the PI.

[0478] II. Not receiving SOC treatment (Not receiving SOC), n = 20 participants: Patients who have not previously received CIDP treatment or patients who received immunoglobulin (IVIg or SCIg) or corticosteroids but stopped treatment for reasons other than lack of response or side effects. Patients should not have received immunoglobulin (IVIg or SCIg) or corticosteroids for at least 6 months prior to screening. Patients have an INCAT score: 2-9 (a score of 2 should be completely from the leg disability component of INCAT).

[0479] III. Successfully treated with SOC therapy (SOC treated), n = 50 participants: Patients receiving stable SOC therapy, defined as no greater than 10% change in frequency or dose of immunoglobulin therapy or corticosteroids in the 8 weeks prior to screening, maintaining stable SOC therapy until the first SAR445088 dose, these patients show documented evidence of an objective and clinically meaningful response, and evidence of clinically meaningful worsening with SOC therapy interruption or dose reduction within 24 months prior to screening. Clinically meaningful improvement is defined as above.

[0480] Because CIDP is a rare disease that limits the number of available study participants, no control group was included in this Phase 2 study, and the study design will allow for an adequate assessment of the safety and efficacy of SAR445088 in this population.

[0481] All participants received SAR445088 at an IV loading dose of 50 mg / kg on day 1, followed by SC injections containing 600 mg SAR445088 per week. Specifically, for the SOC treatment group: after the IV loading dose on day 1, patients treated with IVIg, SCIg or pulse corticosteroids (IV methylprednisolone or oral dexamethasone) started SAR445088 treatment within one week after the last dose of these treatments. Patients who took oral corticosteroids (e.g., prednisone or prednisolone) daily gradually reduced treatment within the first 12 weeks of the study. Therefore, in the first 12 weeks of the study (overlap period), SAR445088 600 mg SC was administered weekly, with an additive effect of SOC treatment.

[0482] Study participants underwent an initial 24-week treatment period (Part A, initial evaluation), followed by an optional extension period that provided up to 52 weeks of additional treatment (Part B). The primary endpoints in this proof-of-concept study were as follows:

[0483] I. SOC-naive and SOC-refractory: Percentage of participants who responded during the SAR445088 treatment period (up to Week 24). Response was defined as a ≥1-point decrease in the adjusted INCAT disability score.

[0484] II. SOC-treated: Percentage of participants who relapsed after withdrawal from the SOC and during the SAR445088 treatment period (up to Week 24). Relapse was defined as an increase in the adjusted INCAT disability score of ≥1 point.

[0485] Secondary objectives of Part A include safety and tolerability, immunogenicity, and efficacy of SAR445088 (using overlapping SOC therapy for the SOC treatment group). The primary objective of Part B is to evaluate the long-term safety and tolerability of SAR445088 in patients with CIDP. The percentage of participants with durable efficacy during the treatment extension period (from Week 24 to Week 76) was defined as no relapse (treated with SOC) or sustained response (SOC refractory and not receiving SOC), defined as no increase of ≥2 points in the adjusted INCAT disability score). Secondary objectives of Part B are the durability of efficacy during long-term treatment, as well as long-term immunogenicity.

[0486] Separate analyses were performed for each patient group based on when each group completed the study. In addition, separate interim analyses were performed when 50% of participants in each group completed the primary 24-week assessment.

[0487] Data from the Phase 2 study will be used to refine the design of a planned Phase 3 study in patients with CIDP.

[0488] Preliminary results

[0489] To date, two pre-planned interim analyses have been conducted in the ongoing Phase 2 study PDY16744. These analyses were conducted when 50% of the participants in the standard of care (SOC) refractory group and 50% of the participants in the SOC treatment group completed 24 weeks, which is the time required to read the primary endpoint. The primary endpoint in the SOC refractory group is the percentage of participants with clinically meaningful responses during the treatment period from day 1 to week 24. Response is defined as a decrease of ≥1 point in the adjusted INCAT disability score relative to baseline. The primary endpoint of the SOC treatment group is the percentage of participants who relapsed (clinically meaningful worsening) during the treatment period from day 1 to week 24. Relapse is defined as an increase of ≥1 point in the adjusted INCAT disability score relative to baseline. INCAT scores range from 0 (normal) to 10 (maximum disability) and represent the sum of the arm and leg scales (5 points each). Patients who experienced a relapse (an increase of 1 point in the adjusted INCAT score relative to baseline) at any time during the first 24 weeks of the study discontinued SAR445088. Results from two interim analyses conducted to date show that:

[0490] · Eight of 12 participants (66.7%) from the SOC refractory group had Clinically meaningful improvement(response, primary endpoint), which was maintained through week 24. The SOC-refractory group consisted of patients with CIDP who had received multiple lines of therapy. Of the 12 SOC-refractory participants included in this analysis, 10 had failed or had an inadequate response to IVIg, 7 of whom had also tried additional therapies (corticosteroids, immunosuppressive therapy, plasma exchange). The remaining 2 participants had previously received corticosteroids and immunosuppressive therapy.

[0491] · Eleven of the 25 participants (44%) from the SOC treatment group switched to SAR445088 after switching from SOC Clinically significant response , which was maintained through week 24. An additional 44% of SOC-treated participants (11 of 25) remained stable (i.e., no change in INCAT disability score) through week 24. The remaining 12% of SOC-treated participants (3 of 25) had a clinically meaningful worsening (relapse, the primary endpoint) after switching from SOC to SAR445088. Of the 25 SOC-treated participants included in the interim analysis, 18 participants were receiving IVIg prior to enrollment, 3 participants were receiving both IVIg and corticosteroid therapy, and 4 participants were receiving corticosteroid therapy alone.

[0492] The clinically meaningful response rates observed in both the SOC-refractory and SOC-treated groups indicate that the dose regimen of SAR445088 tested is effective in CIDP. The results observed in these interim analyses also met the pre-specified criteria for study success and the decision to proceed to Phase 3. Of note, the response rates observed in the SOC-treated group are significant because it was expected that CIDP disease would be controlled in these patients receiving SOC therapy at enrollment, i.e., no improvement was expected.

[0493] Additional clinical efficacy results are shown in Fig.19 and 20 middle. Fig.19 and 20Group data corresponding to changes from baseline in key clinical outcome measures of functional disability (INCAT disability score, I-RODS) and impairment (grip strength, MRC-SS) evaluating the SOC refractory group and the SOC treatment group, respectively, are depicted. The results showed internal consistency, and improvements in each outcome measure followed the expected directionality compatible with a positive effect (consistent decreases in INCAT disability scores over time, combined with consistent increases in I-RODS, grip strength, and MRC-SS over time). The magnitude of the response observed in the analysis indicates a robust effect as it exceeds the minimal clinically important difference (MCID) threshold established in the field. According to consensus, these MCID thresholds are: adjusted INCAT adjusted score: at least 1 point; I-RODS: at least 4 points; grip strength: at least 8-14 kPa, and MRC total score (MRCSS): at least 2-4 points (2021EAN / PNS CIDP Guidelines, Van den Bergh et al. J Peripher Nerv Syst. [Journal of Peripheral Nervous System] 2021; 26(3): 242-268).

[0494] The results of complement biomarker CH50 are shown in Fig.18 The data suggest that the doses tested in PDY16744 (50 mg / kg IV loading followed by 600 mg SC weekly) provide strong and sustained inhibition of complement activity in the blood to a degree (≤10 IU / ml, ≥90% inhibition) that is expected to translate into a beneficial clinical effect, based on extrapolation of the related C1s-targeting compound sultilimab in CAD patients ( et al. Blood. 2019 Feb 28;133(9):893-901).

[0495] Finally, preliminary data on serum neurofilament light chain (NfL) levels ( Fig.21A , 21B ) showed a downward trend over time, which reflects the effects observed in clinical outcomes. NfL is an objective biomarker of axonal damage and has emerged as a potential biomarker of disease activity in autoimmune neuropathies, particularly CIDP (van Lieverloo et al. J Peripher Nerv Syst. 2019 Jun;24(2):187-194.).

[0496] In conclusion, interim data from the ongoing phase 2 study PDY16744 are promising and support that SAR445088 may address an unmet treatment need for patients with CIDP. As a potential limitation of the data, the PDY16744 study was an open-label, uncontrolled study, and therefore it is unclear to what extent a placebo effect could have played a role. However, the response rates observed in this study are significantly better than the placebo rates historically reported in open trials of CIDP, which are in the 11% range for trials using response / improvement as an endpoint (meta-analysis, Lewis et al. J Peripher Nerv Syst. 2020 Sep;25(3):230–237.). This was also true of the 12% relapse rate observed in the SOC treatment group (vs. the historical 43% placebo relapse rate). Furthermore, the combination of the magnitude of the effect, the directionality of each outcome measure, and the correlation with the objective biomarker effect also supports the interpretation that the results are robust and unlikely to be driven by placebo.

[0497] In addition to two interim analyses, preliminary results from the phase 2 study of six patients who had not received SOC treatment showed that the dose regimen tested was effective, with two responses observed among five completers at cutoff. Overall, two patients who had reached week 24 responded to treatment, while one responding patient discontinued the study for reasons unrelated to treatment.

[0498] Additional Proposed Dosage Regimens

[0499] Participants were administered a one-time 50 mg / kg IV loading dose on day 1 and received SC doses administered with a syringe, or pre-filled syringe, or auto-injector, or large-volume drug delivery system. Possible SC doses administered included 300 mgqw, 600 mgqw, 600 mgq2w, 1200 mgq2w, 1200 mgq4w, 2400 mgq4w, 3600 mgq12w, or 7200 mgq12w. Participants received SC doses administered regularly with a syringe, or pre-filled syringe, or auto-injector, or large-volume drug delivery system until the end of treatment.

[0500] Table 13 - Summary of proposed dosage regimens and predicted SAR445088Ctrough during treatment with a large volume subcutaneous device and a 50 mg / kg IV loading dose.

[0501] All references, patents, and patent applications disclosed herein are incorporated by reference with respect to the subject matter in which they are respectively cited, which in some cases may be incorporated in their entirety.

[0502] As used herein in the specification and claims, the indefinite article "a" or "an" should be understood to mean "at least one" unless explicitly stated to the contrary.

[0503] It should also be understood that, in any method claimed herein that includes more than one step or operation, the order of the steps or operations of the method is not necessarily limited to the order in which the steps or operations of the method are described, unless explicitly stated to the contrary.

[0504] In the claims and the foregoing description, all transitional phrases such as "comprising," "including," "with," "having," "containing," "involving," "having," "consisting of," and the like should be understood as open-ended, i.e., meaning including, but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" should be closed or semi-closed transitional phrases, respectively, as set forth in Section 2111.03 of the United States Patent Office Manual of Patent Examining Procedures.

[0505] The terms "about" and "substantially" preceding a numerical value mean ±10% of the stated numerical value.

[0506] Where a range of values ​​is provided, every value between and including the upper and lower ends of that range is specifically contemplated and described herein.

Claims

1. A method for treating cold agglutinin disease in a subject in need thereof, the method comprising administering to the subject about 3.5 g of a humanized antibody that specifically binds complement component C1s, wherein the antibody comprises a light chain (LC) complementary determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HC CDR3 comprising the amino acid sequence of SEQ ID NO:

6.

2. The method of claim 1, wherein the antibody is administered approximately every 10, 12, 14 or 16 weeks.

3. The method of claim 1, wherein the antibody is administered about every 12 weeks.

4. The method of any one of claims 1-3, wherein about 3.5 g of the antibody is administered about 4 weeks after the first dose.

5. The method of any one of claims 1-4, wherein the antibody is administered intravenously or subcutaneously.

6. The method of claim 1, wherein about 3.5 g of the antibody is administered intravenously on day 1 and about every 10, 12, 14, or 16 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29.

7. The method of claim 1, wherein about 3.5 g of the antibody is administered intravenously on day 1 and about every 12 weeks thereafter, and an additional about 3.5 g of the antibody is administered intravenously on or about day 29.

8. The method of any one of claims 1-7, wherein administration of the antibody increases the level of hemoglobin in the subject.

9. The method of any one of claims 1-8, wherein administration of the antibody reduces the level of bilirubin in the subject.

10. The method of any one of claims 1-9, wherein administration of the antibody inhibits classical complement pathway activity by about 90% or more as determined by measuring the amount of terminal complement complex C5b-9.

11. The method of any one of claims 1-10, wherein administration of the antibody reduces the 50% hemolytic complement activity (CH50) level to less than about 10 IU / mL.

12. The method of any one of claims 1-11, wherein after administration of the antibody, the subject has an antibody plasma concentration of at least about 100 μg / mL.

13. The method of any one of claims 1-12, wherein the antibody is contained in a syringe or a vial.

14. The method of any one of claims 1-13, wherein the antibody is administered using a syringe, a pre-filled syringe, or a large volume drug delivery system.

15. The method of any one of claims 1-14, wherein the antibody is administered using an automatic injector.

16. The method of any one of claims 1-15, wherein the antibody comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 7 and a heavy chain variable region (VH), wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:

8.

17. The method of any one of claims 1-16, wherein the antibody is a Fab fragment, a F(ab')2 fragment, a scFv or a Fv.

18. The method of any one of claims 1-16, wherein the antibody comprises a heavy chain constant region of isotype IgG4.

19. The method of claim 18, wherein the IgG4 constant region comprises proline, glutamic acid, leucine and serine substitutions at amino acid residues 108, 115, 308 and 314, respectively, relative to the IgG4 constant region sequence of SEQ ID NO:

11.

20. The method of any one of claims 1-16 and 18-19, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:9 and a heavy chain comprising the amino acid sequence of SEQ ID NO:

10.

21. A method for treating chronic inflammatory demyelinating polyneuropathy (CIDP) in a subject in need thereof, the method comprising administering to the subject a loading dose of about 50 mg / kg of the subject's body weight of a humanized antibody that specifically binds complement component C1s, and one or more maintenance doses of about 300 mg, about 600 mg, about 1200 mg, about 2400 mg, about 3600 mg, or about 7200 mg of the antibody, wherein the antibody comprises a light chain (LC) complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1, a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a LC CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a heavy chain (HC) CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a HC CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HC CDR3 comprising the amino acid sequence of SEQ ID NO:

6.

22. The method of claim 21, wherein the one or more maintenance doses are administered about every 1, 2, 4, or 12 weeks thereafter.

23. The method of claim 21 or 22, wherein the antibody is administered intravenously or subcutaneously.

24. The method of any one of claims 21-23, wherein the loading dose is administered intravenously on day 1, and the maintenance dose is then administered subcutaneously approximately weekly starting on day 8.

25. The method of any one of claims 21-24, wherein the subject is refractory to another CIDP therapy.

26. The method of any one of claims 21-24, wherein the subject is receiving another CIDP treatment within about one week of the loading dose, or is concurrently receiving another CIDP treatment.

27. The method of any one of claims 21-24, wherein the subject has not received another CIDP treatment within about 6 months prior to the loading dose.

28. The method of any one of claims 25-27, wherein the another CIDP treatment is intravenous immunoglobulin (IVIg), subcutaneous immunoglobulin (SCIg), or corticosteroids.

29. The method of any one of claims 19-28, wherein administration of the antibody results in a decrease in the Inflammatory Neuropathy Causes and Treatment (INCAT) disability score by one point or more relative to the INCAT score prior to treatment with the antibody.

30. The method of any one of claims 21-29, wherein after administration of the antibody, the subject has an antibody plasma concentration of at least about 100 μg / mL.

31. The method of any one of claims 21-30, wherein the antibody is contained in a syringe or a vial.

32. The method of any one of claims 21-31, wherein the antibody is administered using a syringe, a pre-filled syringe, or a large volume drug delivery system.

33. The method of any one of claims 21-32, wherein the antibody is administered using an automatic injector.

34. The method of any one of claims 21-33, wherein the antibody comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain variable region (VH), the light chain variable region comprising the amino acid sequence of SEQ ID NO:

8.

35. The method of any one of claims 19-34, wherein the antibody is a Fab fragment, a F(ab')2 fragment, a scFv or a Fv.

36. The method of any one of claims 19-34, wherein the antibody comprises a heavy chain constant region of isotype IgG4.

37. The method of claim 36, wherein the IgG4 constant region comprises proline, glutamic acid, leucine and serine substitutions at amino acid residues 108, 115, 308 and 314, respectively, relative to the IgG4 constant region sequence of SEQ ID NO:

11.

38. The method of any one of claims 21-34 and 36-37, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:9 and a heavy chain comprising the amino acid sequence of SEQ ID NO:10.

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