appendix d pk/pd collection worksheets
appendix c patient's medication diary

| PHASE I RESEARCH URINE SAMPLE COLLECTION SHEET: CYCLE 1 DAY 1                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        |              |                                                                               |                    |
|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------|-------------------------------------------------------------------------------|--------------------|
| CTEP Protocol  P9483                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 | Site:        | Research Nurse:                                                               |                    |
| Dose level:                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          | TRC102 Dose: | Phone:                                                                        | Pager:             |
| Patient ID:                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          | TMZ Dose:    | Page ----------------------------for  sample pick-up  Lab phone: 301-451-1169 | PI: A. P. Chen, MD |
| Phone: 301-768-2749                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  |              |                                                                               |                    |
| PLEASE ENTER DATE, TIME, and VOLUME OF EACH VOID FOR 24 HOURS POST DRUG ADMINISTRATION KEEP SAMPLES REFRIGERATED UNTIL PICKED UP Date Time Volume of  Record comments (i.e., if collection missed), and sign each time  Void you collect a sample Pre-Drug Administration (SAMPLE #100) Administer TRC102 and Temozolomide, Time: DAY 1 Start 24 hour urine collection immediately after study drug administration, keep on ice. Record volume of each void on this sheet  and record total 24 hour volume at the end of the collection. At the end of 24 hours, retain 10 mL aliquot only, place on ice and discard |              |                                                                               |                    |  
| remainder of urine. Charge urine collection container with acid as specified in Appendix E. Time  (start of 24  hour  collection  time) Volume of  Void 24-hour urine collection Record comments (i.e., if collection missed), and sign each time  you collect a sample Volume 1 Volume 2 Volume 3 Volume 4 Volume 5 Volume 6 Volume 7 Volume 8 Volume 9 Total (24-hour  10 mL aliquot obtained (please check box) Sample #101 (10 mL retained from total volume) volume) End 24-hour Urine Collection Time: Date   |
|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|  
Please Send a Copy of this sheet with the last specimen  
| Date:                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         | PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 1 Page -------------------------for  Sample Pick-up Lab phone: 301-451-1169   | Research Nurse:   |                    |        |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|-------------------|--------------------|--------|
| Dose level:                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   | TRC102 Dose:                                                                                                                     | Wt:               | Phone:             | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   | TMZ Dose:                                                                                                                        | BSA:              | PI: A. P. Chen, MD |        |
| *****ONLY PK SAMPLES GO ON ICE******                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          | Phone: 301-768-2749                                                                                                              |                   |                    |        |
| CTEP Protocol P9483                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           | Ht:                                                                                                                              |                   |                    |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal  Actual  Record comments (i.e., if collection missed), and  Time Time sign each time you collect a sample PK 200 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE ----- Day 1 Prior to drug  administration Day 1 Prior to drug  PD 400 4mL NaHep administration Label tube: drug, draw date and time  Administer TMZ and TRC102, Time: PK 201 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE Day 1 1 hour post  dose PK 202 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE Day 1 2 hours post  dose PK 203 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE Day 1 3 hours post  dose PK 204 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE Day 1 4 hours post  dose Continued over |                                                                                                                                  |                   |                    |        |  
| Day                                                                               | Time                                                                                                                    | Instructions                                                                      | Ideal   | Actual   | Record comments (i.e., if collection missed),   |
|-----------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------|---------|----------|-------------------------------------------------|
| Time                                                                              | Time                                                                                                                    | and sign each time you collect a sample                                           |         |          |                                                 |
| PK 205 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE |                                                                                                                         |                                                                                   |         |          |                                                 |
| Day 1                                                                             | 8 hours post  dose                                                                                                      |                                                                                   |         |          |                                                 |
| Day 1                                                                             | 8 hours post                                                                                                            | PD 401 4 mL NaHep                                                                 |         |          |                                                 |
| dose                                                                              | Label tube: drug, draw date and time  PK 206 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE |                                                                                   |         |          |                                                 |
| Day 1                                                                             | 12 hours post  dose                                                                                                     | PK 207 EDTA (lavender top) 2 mL Label tube: drug, draw date and time PLACE ON ICE |         |          |                                                 |
| Day 2                                                                             | 24 hours post  dose                                                                                                     |                                                                                   |         |          |                                                 |  
| Date:                                                                                                                                                                                                                                                                                                                                                                                            | PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 5   |                 |        |        |
|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|-----------------|--------|--------|
| CTEP Protocol P9483                                                                                                                                                                                                                                                                                                                                                                              | Ht:                                                    | Research Nurse: |        |        |
| Page ------------------------- for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                                                                                                                        | PI: A. P. Chen, MD                                     |                 |        |        |
| *****ONLY PK SAMPLES GO ON ICE******                                                                                                                                                                                                                                                                                                                                                             | Phone: 301-768-2749                                    |                 |        |        |
| Dose level:                                                                                                                                                                                                                                                                                                                                                                                      | TRC102 Dose:                                           | Wt:             | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                                                                                                                                      | TMZ Dose:                                              | BSA:            |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal  Actual  Record comments (i.e., if collection missed),  Time Time and sign each time you collect a sample PK 208 EDTA (lavender top) 2 mL Day 5 Prior to dosing Label tube: drug, draw date and time PLACE ON ICE Day 5 Prior to dosing PD 402 4 mL NaHep Label tube: drug, draw date and time |                                                        |                 |        |        |  
| Date:                                                                                                                                                                                                                                                                                    | PHASE I PK/PD SAMPLE COLLECTION SHEET: Cycle 2 Day 1   |                 |        |        |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|-----------------|--------|--------|
| CTEP Protocol P9483                                                                                                                                                                                                                                                                      | Ht:                                                    | Research Nurse: |        |        |
| Page ______________  for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                          | PI: A. P. Chen, MD                                     |                 |        |        |
| *****ONLY PK SAMPLES GO ON ICE******                                                                                                                                                                                                                                                     | Phone: 301-768-2749                                    |                 |        |        |
| Dose level:                                                                                                                                                                                                                                                                              | TRC102 Dose:                                           | Wt:             | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                              | TMZ Dose:                                              | BSA:            |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal Time Actual  Record comments (i.e., if collection missed),  Time and sign each time you collect a sample Day 1 Prior to  PD 403 4 mL NaHep dosing Label tube: drug, draw date and time |                                                        |                 |        |        |
appendix d pk/pd collection worksheets
phase i pk/pd sample collection

| PHASE I PK/PD SAMPLE COLLECTION SHEET: Day 1 each cycle/as  determined by PI Cycle number: ______                                                                                                                                                                                       |                    |                     |        |        |
|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------|---------------------|--------|--------|
| Date:  CTEP Protocol P9483                                                                                                                                                                                                                                                              | Ht:                | Research Nurse:     |        |        |
| Page  for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                                        | PI: A. P. Chen, MD |                     |        |        |
| *****ONLY PK SAMPLES GO ON ICE******                                                                                                                                                                                                                                                    |                    | Phone: 301-768-2749 |        |        |
| Dose level:                                                                                                                                                                                                                                                                             | TRC102 Dose:       | Wt:                 | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                             | TMZ Dose:          | BSA:                |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal Time Actual  Record comments (i.e., if collection missed),  Time and sign each time you collect a sample Day 1 Prior to dosing PD 40X 4 mL NaHep Label tube: drug, draw date and time |                    |                     |        |        |  
| Date:                                                                                                                                                                                                                                                                                                 | PHASE II PD SAMPLE COLLECTION SHEET: Cycle 1 Day 1   |                 |                                                                           |                                         |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------|-----------------|---------------------------------------------------------------------------|-----------------------------------------|
| CTEP Protocol P9483                                                                                                                                                                                                                                                                                   | Ht:                                                  | Research Nurse: |                                                                           |                                         |
| Dose level:                                                                                                                                                                                                                                                                                           | TRC102 Dose:                                         | Wt:             | Phone:                                                                    | Pager:                                  |
| Patient ID:                                                                                                                                                                                                                                                                                           | TMZ Dose:                                            | BSA:            | Page ------------------------- for Sample Pick-up Lab phone: 301-451-1169 | PI: A. P. Chen, MD  Phone: 301-768-2749 |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal  Actual  Record comments (i.e., if collection missed),  Time Time and sign each time you collect a sample Day 1 Prior to drug  400 10 mL Streck administration Label tube: drug, draw date and time |                                                      |                 |                                                                           |                                         |  
| PHASE II PK/PD SAMPLE COLLECTION SHEET: Cycle 1 Day 4 or 5 (day                                                                                                                                                                                                                           |                                          |                 |        |        |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------|-----------------|--------|--------|
| Date:                                                                                                                                                                                                                                                                                     | of biopsy)                               |                 |        |        |
| CTEP Protocol P9483                                                                                                                                                                                                                                                                       | Ht:                                      | Research Nurse: |        |        |
| Page ______________  for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                           | PI: A. P. Chen, MD   Phone: 301-768-2749 |                 |        |        |
| Dose level:                                                                                                                                                                                                                                                                               | TRC102 Dose:                             | Wt:             | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                               | TMZ Dose:                                | BSA:            |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal Time Actual  Record comments (i.e., if collection missed),  Time and sign each time you collect a sample Day 4/5 Prior to  401 10 mL Streck dosing Label tube: drug, draw date and time |                                          |                 |        |        |  
| cycle/as determined by PI Cycle number: ______                                                                                                                                                                                                                                          |                     |                 |        |        |
|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------|-----------------|--------|--------|
| CTEP Protocol P9483                                                                                                                                                                                                                                                                     | Ht:                 | Research Nurse: |        |        |
| Page  for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                                        | PI: A. P. Chen, MD  |                 |        |        |
|                                                                                                                                                                                                                                                                                         | Phone: 301-768-2749 |                 |        |        |
| Dose level:                                                                                                                                                                                                                                                                             | TRC102 Dose:        | Wt:             | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                             | TMZ Dose:           | BSA:            |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal Time Actual  Record comments (i.e., if collection missed),  Time and sign each time you collect a sample Day 1 Prior to dosing 40__ 10 mL Streck Label tube: drug, draw date and time |                     |                 |        |        |  
| PHASE II PK/PD SAMPLE COLLECTION SHEET: Day of restaging followup or progression biopsy                                                                                                                                                                                                                                                                                          |                                          |                 |        |        |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------|-----------------|--------|--------|
| Date:  CTEP Protocol P9483                                                                                                                                                                                                                                                               | Ht:                                      | Research Nurse: |        |        |
| Page ______________  for Sample Pick-up Lab phone: 301-451-1169                                                                                                                                                                                                                          | PI: A. P. Chen, MD   Phone: 301-768-2749 |                 |        |        |
| Dose level:                                                                                                                                                                                                                                                                              | TRC102 Dose:                             | Wt:             | Phone: | Pager: |
| Patient ID:                                                                                                                                                                                                                                                                              | TMZ Dose:                                | BSA:            |        |        |
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Ideal Time Actual  Record comments (i.e., if collection missed),  Time and sign each time you collect a sample Day X Prior to  40__ 10 mL Streck dosing Label tube: drug, draw date and time |                                          |                 |        |        |  
| PHASE II PK/PD SAMPLE COLLECTION SHEET: Optional blood for   |                                                 |                 |        |        |
|--------------------------------------------------------------|-------------------------------------------------|-----------------|--------|--------|
| Date:                                                        | germline WES, collected any time while on trial |                 |        |        |
| CTEP Protocol P9483                                          | Ht:                                             | Research Nurse: |        |        |
| Page ______________  for Sample Pick-up                      |                                                 |                 |        |        |
| Dose level:                                                  | TRC102 Dose:                                    | Wt:             | Phone: | Pager: |
| Patient ID:                                                  | TMZ Dose:                                       | BSA:            |        |        |  
Page ______________ for Sample Pick-up  
Lab phone: 301-451-1169 PI: A. P. Chen, MD  
| Phone: 301-768-2749                                                                                                                                                                                                                                                                |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| PLEASE LABEL EACH TUBE WITH ACTUAL DATE AND TIME OF SAMPLE COLLECTION Day Time Instructions Actual  Actual  Record comments (i.e., if collection missed),  Day Time and sign each time you collect a sample Any day Any time 001 10 mL Streck Label tube: drug, draw date and time |
appendix e pk sample handling
plasma

The 2-mL blood sample is immediately placed on ice and centrifuged within 30 minutes of collection to separate the plasma. The plasma is divided between two tubes, one of which is labeled "A" (which is frozen and stored at -70°C until analyzed) and the other is labeled "B," which contains 25 µL of commercially available 1N hydrochloric acid (HCl): sample "B" is then mixed by vortex action, frozen, and stored at -70°C until analysis.
appendix e pk sample handling
urine

A pre-dose void is collected, its volume measured and recorded, and then a 10-mL aliquot of urine is removed to which 0.4 mL of 1N hydrochloric acid is added, mixed, frozen and stored at -70°C until analysis. The plastic container used for the collection/storage of all voids over the 24-hr collection period is first charged with 25 mL of commercially available 1N HCl prior to the introduction of any patient urine. An additional 25 mL of 1N HCl is added for each 500 mL of urine collected. The contents of the container are mixed by swirling. Each void volume is first measured before adding to the container. At the conclusion of collecting all voids, the contents of the container are mixed by swirling, and then a 10-mL aliquot is removed, frozen, and stored at -70°C until analysis. Send plasma and urine samples to:
Tracy W. Webb Office of the Associate Director/DTP/NCI FNLCR
Boyles Street Building 1047, Room 8 Frederick, MD 21702 301-846-7402 webbtw@mail.nih.gov
appendix f erythrocyte analysis
urine

Please note: As of **Amendment R (5/12/18)***, we will no longer perform this analysis of the* mechanisms of extravascular hemolysis. Blood will be collected for the analysis below in collaboration with the hematology attending consult team. These studies are optional. Blood volumes may be changed at the PI/consult team's discretion. Additional studies (Folate, Vitamin B12, Erythropoietin, TSH) may be done as clinically indicated based on baseline Hgb values. Blood collection tubes, volumes, and service for each test can be found at the Dept. of Transfusion Medicine's Test Guide: http://cclnprod.cc.nih.gov/dlm/testguide.nsf/ When patients develop Hgb level decrement of more than 2 g/dL of baseline during cycle follow-up, contact the hematology consult team for official consultation (Dr. Kazusa Ishii, kazusa.ishii@nih.gov; 301-496-5093); the hematology attending consult team will develop a reference guide/SOP. The hematology consult team will order necessary labs, review the peripheral blood smear, and obtain systematic relevant hematological history at the time of consultation. Laboratory testing/ timing 1) Baseline (within 72 hours prior to enrollment)
CBC with WBC differential Reticulocyte count Peripheral blood smear evaluation LDH LFTs Haptoglobin Ferritin, iron study DAT (Direct Coombs test)
Renal function Urinalysis with microscopy, urine urobilinogen G6PD
Spleen size evaluation (if CT is done recently per protocol, there is no need to repeat. If not, then US spleen)
Osmotic fragility test RBC enzyme evaluation Methemoglobin, venous 2) C1D7 +/- 2 days CBC with diff Reticulocyte count Peripheral blood smear LDH
appendix f erythrocyte analysis
urine

LFTs Haptoglobin DAT Ferritin, iron study Urinalysis with microscopy, urine urobilinogen RBC enzyme evaluation Osmotic fragility test Methemoglobin, venous
appendix f erythrocyte analysis
3) c1d14 +/- 2 days

Reticulocyte count Peripheral blood smear LDH LFTs Haptoglobin DAT
Urinalysis with microscopy, urine urobilinogen