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Table 11: ALPN-101 Flow Cytometric Raw Binding Values (MFI): Mouse Counter Structures
Counter Structure Protein Conc. (pM) ALPN-101 ALPN-101 WT ICOSL-Fc1.1 WT ICOSL-Fc1.1 Fc 1.1 Fc 1.1
CD28 100000 83572 91398 5445 6948 322 300
20000 80814 81971 3961 4032 353 486
4000 89055 93162 3485 3282 284 378
800 25829 25944 3818 3684 245 310
160.0 6174 6235 1879 2070 243 371
32.0 1749 2036 1101 1012 251 314
6.4 1181 885 925 560 328 311
1.3 508 434 322 483 240 348
ICOS 100000 115099 119796 93680 115950 424 540
20000 115509 126740 117357 117763 391 408
4000 99066 86803 84748 91767 328 318
800 42233 41400 43939 37197 378 272
160.0 11003 10863 11799 11055 473 389
32.0 3155 3081 3282 3073 364 10380
6.4 1227 1419 1406 1328 391 1038
1.3 1280 585 1176 1342 548 3710
CTLA-4 100000 43619 49854 2259 1993 280 635
20000 47422 47943 764 786 389 354
4000 40164 51252 639 646 346 378
800 16193 15472 493 664 228 267
160.0 4168 4097 520 616 236 297
32.0 1299 1297 340 330 283 337
6.4 556 643 434 326 324 285
1.3 333 444 457 579 324 764
Pharmacology Study Report Alpine Immune Sciences, Inc. ALPN100-PHRM-001
29
Table 12: ALPN-101 Flow Cytometric Raw Binding Values (MFI): Mock Transfectants
Counter Structure Protein Conc. (pM) ALPN-101 ALPN-101 WT ICOSL-Fc1.1 WT ICOSL-Fc1.1 Fc 1.1 Fc 1.1
100000 6880 5606 2919 4470 297 321
20000 4901 4094 2074 485 194 160
4000 7225 2987 159 281 199 128
800 5994 8204 287 550 132 115
160.0 2671 2370 205 502 166 109
32.0 809 1091 219 239 618 1699
6.4 622 474 142 127 10791 214
1.3 657 188 164 254 184 294
Pharmacology Study Report Alpine Immune Sciences, Inc. ALPN100-PHRM-001
30
Table 13: ALPN-101 BLI Binding Responses for Each Replicate in the Titrations
ICOS CD28 CTLA-4
Conc. (nM) Response Conc. (nM) Response Conc. (nM) Response
144.1 0.769 138.2 0.562 500 0.237
144.1 0.765 138.2 0.626 500 0.269
144.1 0.766 138.2 0.622 500 0.266
72.1 0.551 69.1 0.518 250 0.259
72.1 0.624 69.1 0.514 250 0.276
72.1 0.612 69.1 0.485 250 0.267
36 0.447 34.6 0.426 125 0.228
36 0.479 34.6 0.402 125 0.225
36 0.439 34.6 0.363 125 0.221
18 0.292 17.3 0.299 62.5 0.161
18 0.331 17.3 0.298 62.5 0.168
18 0.323 17.3 0.258 62.5 0.163
9.01 0.166 8.64 0.203 31.3 0.113
9.01 0.215 8.64 0.203 31.3 0.12
9.01 0.208 8.64 0.153 31.3 0.104
4.68 0.087 4.32 0.123 15.6 0.07
4.68 0.123 4.32 0.085 15.6 0.062
4.68 0.141 4.32 0.137 15.6 0.069
2.25 0.08 2.16 0.076 7.81 0.037
2.25 0.074 2.16 0.044 7.81 0.034
2.25 0.075 2.16 0.074 7.81 0.034
Schedule 1.97-C
ALPN-202
ALPN-202 is a homodimeric Fc1.1-fusion protein consisting of 2 identical variant CD80 IgV Fc1.1-fusion protein chains covalently linked through the Fc Hinge region disulfide bonds. Each chain contains a variant CD80 IgV domain fused amino-terminally to the variant human IgG1 Fc designated as Fc1.1.
The amino acid sequence of ALPN-202 is set forth below:
VIHVTKEVKEVATLSCGYNVSVEELEQTRIYWQKDKKMVLTMMSGDLNIWPEYKNRTIFDDITNNLSIMILGLRPSDEGTYECVVLKYEKGAFKREHLAEVTLSVKADGSGGGGSEPKSSDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
Schedule 1.120
Option Exercise Data Package
The Option Exercise Data Package shall include all Clinical Data, including: ALPN-101 Clinical Data, Regulatory Documentation and other Information resulting from the Development Activities or the aGVHD Clinical Study conducted on or prior to (i) in the case of Clinical Information (as defined below), the Option Exerci...
Clinical Studies: All the clinical information for Clinical Study AO3 and AO3a conducted under the Development Plan and the aGVHD Clinical Study (the "Clinical Information"):
o Complete (safety and efficacy) monitored and cleaned Clinical Data set for each patient (SDTM) Clinical protocol, protocol amendments, statistical plan and IB SAS Analysis Data Model (ADaM) data set Report with PK, ADA, and neutralizing antibody (nAb) data Dose recommendation, if applicable Report with Aggregate effi...
Clinical PK, pharmacology and Biomarker Phase study report:
A. Target Saturation (combined CD28/ICOS) B. Change in SLE biomarkers (e.g., total IgM and IgG, Complement C3/C4, ANA and anti-dsDNA) C. Change in peripheral blood numbers and frequencies of T cells, B cells, NK cells and Monocytes (Trucount TBNKMo assay) D. Change in peripheral blood numbers and/or frequency of periph...
NON-CLINICAL 1. Regulatory Documentation
2. Nonclinical and Bioanalytical reports Nonclinical and Bioanalytical reports NHP PK and high concentration SC bridging study reports Bioanalytical methods reports and summaries, including validation reports for quantitation of ALPN-101, ADA and nAb (as appropriate) Standard for Exchange of Nonclinical Data (SEND)-com...
3. CMC Development reports (Cell line, Formulation, Process): Reports for completed activities for cell line development to include: Cell line development including plasmid construction and history of the parental cell line, Clonal cell line generation, Cell line screening and selection, MCB production and characteriza...
Analytical reports Reports including identification and qualification of impurities as needed (process-related or product-related impurities) Reports covering reference standard characterization, qualification and stability Report(s) documenting characterization results to include physicochemical, biophysical, and biol...
All CRO agreements, data and reports and GxP quality agreements and audits.
Schedule 1.121
Option Exercise Data Package Trigger Event
The Option Exercise Data Package Trigger Event shall occur upon the first date on which all of the following conditions are satisfied:
CLINICAL (a) the Phase IIa Clinical Study, initiating with a Week 12 endpoint (AO3) with the option to amend to a Week 24 endpoint (AO3a) once the 6-month GLP tox study is available, has the minimum number ("n") of Response-Evaluable Patients identified below for each arm of such Clinical Study and are otherwise conduc...
(b) a complete set of all Clinical Information that includes such Information generated during the First Cohort (with the Week 12 endpoint (AO3)) after enrollment of the last Response-Evaluable Patient enrolled across First Cohort and if amended, the Second Cohort (with the Week 24 endpoint (AO3a)) is available to Lice...
As used herein, "Response-Evaluable Patient" means a patient enrolled in either an arm or Cohort of the Phase IIa Clinical Study, as applicable, that has completed a 12-week cycle of treatment in accordance with the study protocol for such Clinical Study (as reflected in the Development Plan).
As used herein, the First Cohort and (if amended) the Second Cohort of the Clinical Study shall include the following elements: • First Cohort of Phase IIa - (Week12 endpoint (AO3)) – ALPN-101 monotherapy IV in SLE (max n=130)
* Final design to be mutually agreed upon by the Joint Governance Committee in writing prior to study initiation
• Second Cohort of Phase IIa (amended to week 24 endpoint (AO3a)) – ALPN-101 monotherapy SC in SLE (max n=130)
Schedule 1.157
Bridging Study Success Completion Criteria
Objective: Evaluate suitability of the new high concentration SC formulation to replace the initial formulation used in the 12-Week Phase IIa SLE Clinical Study.
Successful bridging will be established based on: (1) relative bioavailability estimates for Cmax and AUC of the IV formulation in the human PK study; (2) comparable immunogenicity profile across the two formulations; (3) acceptable safety in terms of injection-site reactions; and (4) achievement of the criteria set fo...
Licensor may commence the Bridging Study following the completion of the pre-formulation/formulation Development Activities set forth in the Development Plan with a new high concentration (>70mg/mL) SC formulation which meets the following criteria:
Target volume < 1 mL per single dose with dosing no more frequently than weekly Needle size not larger than G27 Max. injection force at 2-8°C not more than 20N, measured at 80 mm/min
At least 3-month drug product stability data with the development batch in representative container closure system o Acceptable stability profile defined as Recommended long term storage condition 2—8°C Comparable stability profile to current 25 mg/mL vial (e.g. main peak icIEF > 30%, HMW (SEC) < 5% at 2-8°C)
Schedule 2.1.1
Initial Development Plan
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