IL-17 antibody formulations and methods of treatment using same

Abstract

The present invention provides stabilized pharmaceutical formulations for anti-IL-17 antibodies, comprising e.g. citrate, sodium chloride and polysorbate-80 at pH 5.7. These stabilized anti-IL-17 antibody pharmaceutical formulations can be used to treat rheumatoid arthritis, psoriasis, ankylosing spondilitis, psoriatic arthritis or multiple myeloma.

Claims

1. A pharmaceutical formulation comprising an anti-IL-17 antibody at a concentration in the range of about 80 mg/ml to about 150 mg/ml, citrate buffer at a concentration of about 20 mM, sodium chloride at a concentration of about 200 mM, polysorbate-80 at a concentration in the range of about 0.02% (w/v) to about 0.03% (w/v), and pH at about 5.7, wherein the anti-IL-17 antibody comprises a light chain variable region given by the amino acid sequence of SEQ ID NO:2, and a heavy chain variable region given by the amino acid sequence of SEQ ID NO:3.

2. The formulation of claim 1, wherein the concentration of the anti-IL-17 antibody is about 80 mg/ml.

3. The formulation of claim 2, wherein the formulation is an anti-IL-17 antibody pharmaceutical solution formulation.

4. A method of treating rheumatoid arthritis, psoriasis, ankylosing spondilitis, psoriatic arthritis or multiple myeloma comprising administering to a patient in need thereof an effective amount of the pharmaceutical formulation of claim 1.

5. A method of treating rheumatoid arthritis, psoriasis, ankylosing spondilitis, psoriatic arthritis or multiple myeloma comprising administering to a patient in need thereof an effective amount of the pharmaceutical formulation of claim 2.

6. A method of treating rheumatoid arthritis, psoriasis, ankylosing spondilitis, psoriatic arthritis or multiple myeloma comprising administering to a patient in need thereof an effective amount of the pharmaceutical solution formulation of claim 3.

Description

EXAMPLE 1

(1) Formulating mAb 126

(2) TABLE-US-00001 TABLE 1 mAb 126 Drug Product Formulation Concentration Component (mg/mL) mAb 126 80 Sodium Citrate Dihydrate 5.106 Citric Acid Anhydrous 0.507 Sodium Chloride 11.69 Polysorbate 80 0.30 Water for Injection q.s. to 1 mL Hydrochloric Acid pH adjustment Sodium Hydroxide pH adjustment

(3) TABLE-US-00002 TABLE 2 Buffer Excipient Composition Component mg/mL Sodium Citrate Dihydrate 5.106 Citric Acid Anhydrous 0.507 Sodium Chloride 11.69 Polysorbate 80 0.414

(4) The manufacturing process for the anti-IL-17 antibody pharmaceutical solution formulation for mAb 126 (Table 1) consists of first compounding of the buffer excipient composition (Table 2), followed by compounding of the final drug product formulation.

(5) The buffer excipient composition (Table 2) is prepared, filtered, and stored for drug product formulation compounding. An appropriate quantity of water at a temperature of 20+/−5° C. is weighed into a tared empty vessel of appropriate size. The appropriate quantity of sodium citrate is added and mixed; the appropriate quantity of citric acid is then added and mixed. Then, the appropriate quantity of sodium chloride is added and mixed. Polysorbate 80 is accurately weighed out in a glass container and an appropriate quantity of water at a temperature of 20+/−5° C. is added into the glass container to give the concentration in Table 2, and the solution is mixed. The entire content of the polysorbate 80 solution is added to the other excipients. The polysorbate 80 solution container is rinsed with water to ensure the entire content is transferred. After addition of the polysorbate 80 solution, the solution is mixed. After dissolution and mixing has been completed, the pH of the solution is checked to be within 5.7+/−0.2; adjustment with HCl or NaOH solution is done if necessary. The buffer excipient composition is passed through a fiber (polyvinylidene fluoride [PVDF]) for bioburden reduction.

(6) The buffer excipient composition is supplemented with additional polysorbate 80 to account for the lower concentration (0.2 mg/mL polysorbate-80) in the active pharmaceutical ingredient (API) compared to the final drug product formulation. Since the concentration of mAb 126 in the API will vary, the amount of buffer needed for dilution will also change. This variation requires that the concentration of polysorbate 80 be adjusted for the buffer excipient composition recipe with each batch.

(7) The stored mAb 126 API containers (mAb 126 is expressed in cells, purified, and concentrated; the resulting API is then frozen at about 150 mg/mL to about 160 mg/mL mAb 126 in 20 mM citrate buffer, 200 mM NaCl, 0.02% polysorbate-80, at pH about 5.7) are equilibrated to a temperature of 20+/−5° C. The API solution is then mixed with an appropriate amount of the buffer excipient solution to achieve 80% of the theoretical batch size. The pH of the solution is checked to be within 5.7+/−0.2. After pH adjustment, the solution is mixed and a sample is taken for an in-process UV assay to determine the mAb 126 concentration. An appropriate quantity of the buffer excipient solution is added to reach the final target batch weight. After mixing, the pH of the solution is checked to be within 5.7+/−0.2, and the osmolality of the solution is checked to be within 360-480 mOsm/Kg. The mAb 126 drug product solution is passed through PVDF filter for bioburden reduction and stored at 5° C.

EXAMPLE 2

(8) Phase Separation

(9) mAb 126 is found to have a propensity to phase separate while in solutions that are below 0° C., Liquid-liquid phase separation (LLPS) needs to be solved since storage of the mAb 126 drug product will be at 5° C. Storage of drug product at 5° C. requires stability for periodic refrigerator temperature excursions below 0° C. Increasing the NaCl concentration is shown to lower the temperature at which LLPS occurs in mAb 126 formulations, and increasing the citrate concentration is also shown to lower the temperature at which LLPS occurs in mAb 126 formulations.

(10) LLPS events are tested based on a technique developed specifically for the temperature range (phase separation occurring between −12 and 0° C.) encountered for mAb 126. Two milliliter (mL) samples of 10 to 200 mg/mL mAb 126 were placed in a LyoStar II (FTS Systems) lyophilization unit with the cycle shown in Table 3. Pressure was kept at atmospheric for the experiments. Sample conditions are tested at least three times and the standard deviation is approximately 0.5° C. for samples measured in triplicate. The samples are visually monitored during the lyophilization cycle for signs of phase separation, including cloud point (white opaque appearance) and the formation of a dense, protein rich layer at the bottom of the vial. Samples appear increasingly opalescent during cooling, but this effect is easily differentiable from cloud point. When cloud point occurs, the sample becomes an opaque white solution in less than a second versus a gradual increased opalescence where an object behind a vial is still visible through the sample.

(11) TABLE-US-00003 TABLE 3 Lyophilization Cycle for Liquid-Liquid Phase Separation Testing Step 1 2 3 Cooling Rate 5 1 5 (° C./min) Target (° C.) 5 −30 20 Hold Time (min) 10 20 >20

(12) As shown in Table 4, increasing the NaCl concentration is shown to depress the cloud point temperature. The factors which limit increasing the NaCl concentration in the anti-IL-17 antibody pharmaceutical formulation are hyper-tonicity and other non-phase-separation effects; 200 mM NaCl avoids these problems while still depressing the cloud point temperature. Also, LLPS mAb126 is shown to be bimodal at all NaCl concentrations, except 300 mM NaCl; LLPS is strongest at a concentration of 100 mg/mL mAb 126, and the phase separation effects lessen as the concentration moves higher or lower than 100 mg/mL.

(13) TABLE-US-00004 TABLE 4 Liquid-Liquid Phase Separation: NaCl and mAb 126 Concentration Effects (Cloud Point ° C.) mAb 126 Conc. (mg/mL) 10 50 100 150 200 50 mM NaCl −4.1 2.1 5 100 mM NaCl −1.2 2.7 1.1 150 mM NaCl −7.5 −4 −5.7 −10.4 200 mM NaCl −8.5 −7 −10.2 −10.8 250 mM NaCl −6.8 −11.3 300 mM NaCl −12.5 −9.9 −12.4 −8.8

(14) The effect of pH on cloud point of a mAb 126 solution is measured. The cloud point was found to be minimized at two pHs, pH 4 and pH6. pH 6 should be considered optimal over pH 4 when selecting a formulation for mAb 126, because chemical instability of mAb 126 at pH 4 precludes using the low cloud point at 4. Given the gel formation at pH 6.3-6.4, shown in Example 3, for the mAb 126 pharmaceutical formulations, pH 5.7+/−0.3 is preferred over pH 6+/−0.3 in order to allow for a pH range that remains stable over the shelf-life of the pharmaceutical formulation.

(15) Effects of various commonly used buffers on liquid-liquid phase separation are explored to understand what the optimal buffer system would be for mAb 126 (Table 5); citrate buffer is found to be most effective at reducing the temperature at which LLPS occurs. These studies are conducted with 150 NaCl. Acetate buffer is comparable to citrate buffer at pH 5 in terms of the effect on cloud point; however, chemical instability at pH 5 makes this pH condition less favorable.

(16) The concentration of citrate is shown to positively affect LLPS (Table 6). While citrate depresses cloud point up to 50 mM; citrate buffer is also reported to cause more pain on injection. As such, citrate concentrations higher than 30 mM are likely to be unacceptable from a patient compliance standpoint.

(17) TABLE-US-00005 TABLE 5 Buffer Effects on Cloud Point Temperature with 150 mg/mL mAb 126 Cloud Point Buffer mM pH (° C.) Acetate 10 5 −4.1 Citrate 10 5 −4.4 Histidine 10 6 −2.2 Citrate 10 6 −8.1

(18) TABLE-US-00006 TABLE 6 Liquid-Liquid Phase Separation: Citrate Concentration Effects with 150 mg/mL mAb 126 Citrate conc. Cloud Point (° C.)  5 mM −6.3 10 mM −8.2 20 mM −10.2 30 mM −13.6 40 mM −14.6

EXAMPLE 3

(19) Gel Formation

(20) Biologic drug products are stored at 5° C.+/−3° C. to minimize chemical and physical degradation over the shelf-life of a product. Events such as thermodynamic solid phase change or gel formation are typically not acceptable, even if reversible, because they can negatively affect stability and hinder the required visual inspection of samples prior to use.

(21) Solid phase change is observed in high concentration samples of mAb 126 at pH conditions <pH 5 and >pH 7 at 5° C. This thermodynamic event is shown to be reversible by equilibrating the vials at room temperature. Consequently, samples are tested at various pH conditions and monitored for thermodynamic changes in order to find the phase boundaries more precisely. These tests are conducted by dialyzing samples into pH 7, citrate buffer at 5° C. to induce phase change. The solid material is then dialyzed at the same temperature into conditions of interest to ascertain if samples would reverse to the solution state. Equilibrium testing method is preferable to long-term sample storage with intermittent inspections because, while a particular formulation may be thermodynamically unstable, the kinetics of solid phase change may require months to occur in some cases.

(22) Studies are conducted with 100 mg/mL or 150 mg/mL mAb 126 at 200 mM NaCl, 10 mM citrate buffer in 0.1 pH increments to find where the boundaries are for solid phase formation. The transition between phases is found between pH 6.3 and 6.4 at 5° C. Because of the phase boundary, the target pH for a mAb 126 formulation is lowered from pH 6 to 5.7 to ensure a stable storage window.

(23) Two experiments with 80 mAb 126 at 5° C. are carried out in 20 mM citrate buffer, 200 mM NaCl, and 0.03% polysorbate-80; there is no gel formation at or below pH 6.1, while gel formation occurs above pH 6.1. These experiments indicate that the formulation with 80 mg/mL mAb 126 has a window from pH 5.7+/−0.3 pH units wide that avoids gel formation.

EXAMPLE 4

(24) Chemical Instability

(25) For a pharmaceutical formulation to achieve stability, both physical and chemical sources of instability need to be addressed in the formulation. Chemical instability can result in degradation of the antibody.

(26) In order to assess the effect of pH on chemical stability of mAb 126 at 100 and 150 mg/mL, samples of mAb 126 are analyzed for increases in total % acid variants by chromatography. The range of pH 4 to 7 is explored in half pH unit increments. The buffer for the study includes 10 mM citrate buffer, 150 mM NaCl and 0.02% polysorbate 80. Two mL solutions are stored in 3 glass vials with serum stoppers, Samples in these pH environments are stored at 5° C., 25° C. and 40° C. in order to more accurately model the temperature effect on the different forms of degradation. Samples are analyzed using Cation Exchange (CEX) HPLC using UV detector and a Dionex ProPac WCX-10 column (4×250 mm) using pH 6 10 mM Bis Tris Propane (Mobile Phase A) and pH 9.6 10 mM Bis Tris Propane, 50 mM NaCl (Mobile phase B).

(27) By CEX, increases in total % acid variants (% AV) are the more reliable indicator of degradation for mAb 126. Vials of mAb 126 stored at 25° C. are shown by CEX to be most stable between pH 5-6 and least stable at more alkaline conditions. When samples are stored at 40° C., pH 5 and 5.5 are the most stable, with 6 appearing only slightly more stable than the higher pH environments. These results show that the pH of the pharmaceutical solution formulation for mAb 126 should be between pH 5 and pH 6.

EXAMPLE 5

(28) Design of Experiment (DOE) Study

(29) The DOE study uses a multivariate approach to examine the physical and chemical stability of mAb 126 solution formulations. mAb 126 solution formulations are prepared according to Table 7. Each variable is explored at five levels to measure any curvature which might exist in the output response parameters or interactions between the input variables. The center point condition for the experiment is 20 mM citrate, 200 mM NaCl, pH 5.7, 0.02% polysorbate 80. Three center points are dispersed throughout the design. Independently preparing three center points provides an estimation of certainty in the analytical data without requiring all of the conditions to be prepared and analyzed in duplicate or triplicate. Samples are stored at four temperature conditions (5, 25, 30 and 40° C.). This range of temperatures allows for estimations of the activation energies for the results. Additionally, higher temperature storage enables earlier predictions of optimal formulation conditions.

(30) A number of analytical techniques are selected to monitor chemical and physical stability including size exclusion chromatography (SEC), cation exchange chromatography (CEX) HPLC, HIAC-based particle analysis, digital-image based particle analysis by micro-flow imaging (MFI, Protein Simple/Brightwell Model DPA 4200 with size range of 2-100 um), visual appearance, reduced and non-reduced Bioanalyzer Lab-on-a-Chip (LoC), pH, viscosity, and UV absorption (to measure protein content).

(31) Using the data from all temperatures from the initial three month period, activation energies (Ea) are calculated employing an Arrhenius kinetic model (zero or first Order). Energies are found using non-linear regression of all Runs. The model is then employed to extrapolate trends out to 24 months at the relevant commercial storage temperature (5° C.). Zero order Arrhenius modeling is used for SEC (monomer, polymer Rel. Sub/Impurities), CEX (acid variants), reduced/non-reduced LoC, and UV content. The best fit for CEX basic variant trends is a first order model.

(32) TABLE-US-00007 TABLE 7 Experimental Design Run pH [PS80[ [NaCl[ [mAb 126[ [Buffer[ Buffer Type 1 5.7 0.02 200 120 20 Citrate 2 6 0.01 150 105 25 Citrate 3 6 0.03 150 105 15 Citrate 4 5.7 0 200 120 20 Citrate 5 5.4 0.03 150 135 15 Citrate 6 6 0.01 250 105 15 Citrate 7 5.7 0.02 200 90 20 Citrate 8 5.7 0.02 100 120 20 Citrate 9 6.3 0.02 200 120 20 Citrate 10 5.1 0.02 200 120 20 Citrate 11 6 0.03 250 105 25 Citrate 12 5.4 0.01 150 105 15 Citrate 13 5.4 0.03 250 135 25 Citrate 14 6 0.03 150 135 25 Citrate 15 5.7 0.02 200 120 20 Citrate 16 5.7 0.02 200 120 10 Citrate 17 5.4 0.01 250 135 15 Citrate 18 5.4 0.01 150 135 25 Citrate 19 6 0.03 250 135 15 Citrate 20 5.7 0.04 200 120 20 Citrate 21 5.4 0.03 250 105 15 Citrate 22 6 0.01 150 135 15 Citrate 23 5.4 0.03 150 105 25 Citrate 24 5.7 0.02 300 120 20 Citrate 25 5.7 0.02 200 150 20 Citrate 26 6 0.01 250 135 25 Citrate 27 5.4 0.01 250 105 25 Citrate 28 5.7 0.02 200 120 30 Citrate 29 5.7 0.02 200 120 20 Citrate
Site Exclusion Chromatography

(33) The two variables with the strongest effect on percent monomer based on SEC results are pH and mAb 126 concentration. The effect of mAb 126 concentration is linear with increasing protein concentration resulting in decreased monomer purity.

(34) Three input variables (pH, NaCl concentration, and buffer concentration) show curvature in their effect on percent monomer. With respect to NaCl and citrate concentration, values near the center point are the most stable. Lower pH conditions are slightly more stable than the center point, but other degradation pathway results make lowering the target pH less attractive.

(35) Interactions between the effect of pH and protein concentration occur with the effects of protein concentration being reduced at pH conditions below 5.7. The two year prediction for monomer purity at the center point conditions of the study is slightly below 96.7%. Polysorbate 80 concentration has little effect on stability from 0.02-0.04%.

(36) Cation Exchange Chromatography

(37) The CEX analysis focuses on the growth of acidic species over time. Statistical modeling of CEX % AV (Ea is 23.7 kcal/g-mol) shows that little chemical modification should be expected after 24 months of storage at 5° C. Acidic variant generation is minimized near the center point for pH, but increases with greater citrate concentration. mAb 126 and Polysorbate 80 concentration trends suggest that the center point is close to the least optimal position; however, based on the magnitude of the y-axis scale, the difference in center point stability to other conditions is essentially negligible.

(38) Reduced Bioanalyzer LoC

(39) Reduced LoC percent purity is a combination of the relative percentages of heavy and light chains. The two input variables with the strongest influence on the 24-month predictions are pH and NaCl concentration. Percent purity is maximized near the center point of 200 mM NaCl. The percent purity increases with increasing pH. Even at the extreme formulation conditions tested in the DOE study, the molecule is still >98% pure, indicating that the antibody is stable as measured by reduced LoC over the range of the multivariate analysis. The Ea is 21.8 kcal/g-mol.

(40) Combined Projections

(41) All conditions in the design space studied during this experiment have 2 year shelf life projections that predict <5% degradation. However, other physical factors preclude pH conditions above 6.3; therefore, the target conditions for the formulation should not be near this pH edge. Additionally, it is important to select a target that is in an optimal global maximum for the input variables explored. Based on manufacturing considerations, which indicate that >0.01% polysorbate 80 is needed for pumping, the polysorbate 80 target is 0.03%. Based on the results of this study, the optimal formulation conditions is 20 mM citrate, 200 mM NaCl, pH 5.7 with 0.03% polysorbate 80.

EXAMPLE 6

(42) Stability at 80 mg/mL mAb 126

(43) The stability of 80 mg/mL mAb 126 in 20 mM citrate, 200 mM NaCl, pH 5.7 with 0.03% polysorbate 80 is tested out to 24 months. For storage at 5° C., stability of the anti-IL-17 antibody pharmaceutical formulation is measured at 0, 1, 3, 6, 9, 12, 18, and 24 months. 5° C. will be the expected storage temperature for the anti-IL-17 antibody pharmaceutical formulation. Accelerated stability studies at 25 are run for 1, 3, and 6 months.

(44) A number of analytical techniques are selected to monitor chemical and physical stability including size exclusion chromatography HPLC, cation exchange chromatography HPLC, visual appearance, pH, and UV absorption. CE-SDS is performed utilizing the Beckman Coulter IgG Purity/Heterogeneity kit with a Beckman Coulter ProteomeLab PA800 Enhanced or Plus capillary electrophoresis (CE) instrument. For Reduced CE-SDS, samples are analyzed in a bare-fused silica capillary under denatured, reducing conditions by molecular sieving through a replaceable gel polymer matrix after each sample injection. For non-reduced CE-SDS, samples are diluted to approximately 5 mg/mL in water and subsequently diluted in sample diluent (20 mM IAM in 100 mM Tris, 1% SDS, pH 9.0) to approximately 1 mg/mL. Afterwards samples are analyzed in a bare-fused silica capillary under denatured, non-reducing conditions by molecular sieving through a replaceable gel polymer matrix at constant voltage. For both methods, UV detection is performed at 214 nm. Results are shown in Table 8.

(45) TABLE-US-00008 TABLE 8 Stability Data for mAb 126 at 80 mg/mL Analytical Storage Month Property Condition 0 1 3 6 9 Potency 5° C. 87 — — — 108   (Bioassay), % 25° C./60% 106   — 109 — RH Quantity (UV), 5° C. 75.4 — 76.2 75.5 75.4 mg/mL 25° C./60% 75.3 76.0 76.5 — RH Monomer 5° C. 98.3 — 98.1 98.3 97.9 Purity (SEC), % 25° C./60% 98.3 97.6 97.6 — RH Rel Subs/ 5° C. 1.7 — 1.9 1.7  2.1 Impurities: 25° C./60%  1.7 2.4 2.4 — Total (SEC), % RH mAb 126 5° C. 97.8 — 97.4 97.3 97.2 Purity (CE-SDS, 25° C./60% 97.4 96.9 96.3 — Reduced), % RH Charge 5° C. 53.9 — 53.0 54.1 55.1 Heterogeneity 25° C./60% 56.7 59.7 60.0 — (CEX) [Main RH Peak], % Charge 5° C. 13.4 — 15.4 15.3 16   Heterogeneity 25° C./60% 15.8 21.4 27.4 — (CEX) [Acidic RH Variants], % Charge 5° C. 32.8 — 31.6 30.7 28.9 Heterogeneity 25° C./60% 27.5 18.8 13.0 — (CEX) [Basic RH Variants], % pH 5° C. 5.7 — 5.7 5.7  5.7 25° C./60%  5.7 5.7 5.7 — RH Physical 5° C. NT Pass.sup.1 — — Pass Appearance 25° C./60% NT — — — RH Particulate 5° C. 289 — 131 184 63   Matter 25° C./60% 194   177 369 — (greater than RH or equal to 10 micrometer), particles per container Particulate 5° C. 9 — 8 31 3  Matter 25° C./60% 4  60 20 — (greater than RH or equal to 25 micrometer), particles per container .sup.1Physical appearance was not performed at lot release or the 1 month time point. Result shown was obtained after storage at 5° C. for approximately 2.5 months.

(46) TABLE-US-00009 Sequence Listing (human IL-17) SEQ ID NO: 1 MTPGKTSLVS LLLLLSLEAI VKAGITIPRN PGCPNSEDKN FPRTVMVNLN IHNRNTNTNP KRSSDYYNRS TSPWNLHRNE DPERYPSVIW EAKCRHLGCI NADGNVDYHM NSVPIQQEIL VLRREPPHCP NSFRLEKILV SVGCTCVTPI VHHVA (LCVR) SEQ ID NO: 2 DIVMTQTPLS LSVTPGQPAS ISCRSSRSLV HSRGNTYLHW YLQKPGQSPQ LLIYKVSNRF IGVPDRFSGS GSGTDFTLKI SRVEAEDVGV YYCSQSTHLP FTFGQGTKLE IK (HCVR) SEQ ID NO: 3 QVQLVQSGAE VKKPGSSVKV SCKASCYSFT DYHIHWVRQA PGQGLEWMGV INPMYGTTDY NQRFKGRVTI TADESTSTAY MELSSLRSED TAVYYCARYD YFTGTGVYWG QGTLVTVSS (Light chain) SEQ ID NO: 4 DIVMTQTPLS LSVTPGQPAS ISCRSSRSLV HSRGNTYLHW YLQKPGQSPQ LLIYKVSNRF IGVPDRFSGS GSGTDFTLKI SRVEAEDVGV YYCSQSTHLP FTFGQGTKLE IKRTVAAPSV FIFPPSDEQL KSGTASVVCL LNNFYPREAK VQWKVDNALQ SGNSQESVTE QDSKDSTYSL SSTLTLSKAD YEKHKVYACE VTHQGLSSPV TKSFNRGEC (Heavy chain) SEQ ID NO: 5 QVQLVQSGAE VKKPGSSVKV SCKASGYSFT DYHIHWVRQA PGQGLEWMGV INPMYGTTDY NQRFKGRVTI TADESTSTAY MELSSLRSED TAVYYCARYD YFTGTGVYMG QGTLVTVSSA STKGPSVFPL APCSRSTSES TAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTKTY TCNVDHKPSN TKVDKRVESK YGPPCPPCPA PEFLGGPSVF LFPPKPKDTL MISRTPEVTC VVVDVSQEDP EVQFNWYVDG VEVHNAKTKP REEQFNSTYR VVSVLTVLHQ DWLNGKEYKC KVSNKGLPSS IEKTISKAKG QPREPCVYTL PPSQEEMTKN QVSLTCLVKG FYPSDIAVEW ESNGQPENNY KTTPPVLDSD GSFFLYSRLT VDKSRWQEGN VFSCSVMHEA LHNHYTQKSL SLSLG