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PF-07302048RatPharmacokinetics
PF-07302048_06Jul20_07242 4
PFIZERCONFIDENTIAL
Page 1of 15A SINGLE DOSE PHARMA COKINETICS STUDY OF ALC-0315 AND ALC -0159
FOLLOWING INTRAVENOU S BOLUS INJECTION OF PF-07302048
NANOPARTICLE FORM ULATION IN WISTAR HA N RATS
This document contains co nfidential information belonging to Pfizer. Except as may be otherwise agreed to in
writing, by accepting or reviewing these materials, you agree to hold such information in confidence and not to
disclose it t o others (except where required by applicable la w), nor to use it fo runauthorized purposes. In the
event of actual or suspected breach of this obligation, Pfizer should be promptly notified.
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Page 2of 151.SUMMAR Y
Following a single IVbolus administration of a luciferase -encoding mod RNA formulated in
anLNP with an identical lipid composition as PF -07302048 ( SARS-CoV-2 mRNA Vaccine ;
BioNTech code num ber BNT162) at 1mg/kgto maleWistar Han rats, plasma concentrations
of ALC-0315and ALC-0159(the novel excipients in P F-07302048 )decreased rapidly,with
initial t½of1.62 and 1.72 h, respectivel y. ALC-0315and ALC-0159 were then cleared from
plasma, resulting in terminal elimination t½of 139and 72.7 h, resp ectively.
Theestimated percent of dose distributed to the l iverwas~60% for AL C-0315 and ~ 20% for
ALC-0159. The percent of dose excreted u nchanged infeceswas ~1% for AL C-0315 and
~50%for ALC-0159. The percent of dose excr eted unchanged in the urin ewas not
calculated due to all values being BLQ.
2.OBJECTIVE
The PF-07302048 vaccine LNPformulation contains twonovel excipients, ALC-0315 and
ALC-0159,inthe nanopa rticle.The objec tive of this study is to assess the pharmacokinetics
and elimination of ALC-0315 andALC-0159 following a single IVbolus administration of a
luciferase -encoding mod RNA with an identical na noparticle lipid composition as
PF-07302048at 1mg/kg to male Wist arHanrats.
3.MATERIALS AND METHODS
3.1. Preparation of Doses
IV
Study ID PF-07302048_06Jul20_072424
Compound lotnumber FM-1261-A
Dose of modRNA(mg/kg) 1(1.96mg/kg ALC-0159, 15.3mg/kg ALC -0315)
Formulation concentration (m g/mL) 1
Dose volume (mL /kg) 1
3.2.StudyConduct
IV
In-life location Pfizera
Species (strain) Rat (Wistar Han)
Sex/number of animals Male/3 animals per time pointb
Feeding Condition Fasted
Administration type IV bolus
Administration site Lateral tailvein
Sampling site Inferior vena cava
Blood and liver samplingtime points (hpostdose) Predose, 0.1, 0.25 , 0.5, 1, 3, 6,24,48,96, 192,
336
Anticoagulant K2EDTA
Urine and feces sample collection i nterval(hpostdose)cPredose(-24-0), 0-24, 24-48,48-72,72-96,
96-120,120-144,144-168, 168-192, 192-216,
216-240, 240-264, 264-288,288-312, 312-336
a. Pfizer Worl dwide Research, Deve lopment and Medical ,San Diego , CA.
b. Non-serial sampling, 36 animals total.
c. Urine and fece s were collected from animals34-36,placed in metabolism cages.
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Page 3of 153.3.Bioanalyt ical Summary for Qua ntitation of ALC -0315 and ALC -0159in Plasma,
LiverHomogenates, Urine , and Feces Homogenates
Bioanalytical Platform (instrument) LC-MS/MS(AB Sciex QTRAP 5500 )
Mobile phase A:0.1% formic acidwith 10 mM ammonium format e
B:Acetonitrile wi th 10mM ammoni um formateand0.1% formic acid
Flow rate 0.25 mL/min
Gradient Time (min) Mobile Ph ase A (%)Mobile Phase B (%)
0.1 0 100
0.5 0 100
2.5 55 45
4.2 55 45
4.5 0 100
6.5 0 100
Column WatersAtlantis HILIC Silica 2.1 ×100mm, 3µ
Detection mode Positiveselected reaction monitoring mode
Datacollection software/ version Analyst Version 1.7.1
Data analysi s software/ versi on Watson Version 7.5
MRM tra nsitions 766.9510.7 (ALC-0315)
839.2494.7(ALC-0159)
837.1264.6(PEG-2000, ISTD)
Additionalconditions can be found i n the analysis files referenced in Section 5(Archiving) .
3.4.Method Summary
1.In brief, 20L aliquots of plasma, liver homoge nate(sections from 3 areas in theliver
were homogenized, p ooled, and diluted 1:20 or 1:100 with blank sample matrix when
necessary ), urine, and feces homogenate (diluted 1:10 or 1:20with blank sample matrix
when necessary ) samples and standardswere subje ctedto proteinprecipitatio n with
acetonitrile containing an internal st andard, PEG-2000. Samples were vortexed and
centrifuged to obtain supernatant,which was analyzed using LC-MS/MS.
2.Method informa tion was recorded and a rchival reco rds are available as described in
Section 5(Archiving).
3.Analystwas used to measure p eak areas an d peak area ratios of anal yte tointernal
standard were ca lculated. A calibration curve was constructed from the pe ak area rati os
(analyte to internalstandard) with aquadratic(1/x)regression using Watson LIMS. The
linear dynamic range s of the standard cu rvesfor ALC-0315 and ALC -0159were4.88to
2500 ng/mLfor plasma, 19.53 to 10000 ng/g for liver, 4.88 to 2500 ng/mLfor urine, and
6.592 to3375 ng/g for feces.
3.5. Data Analysis
Generation and analyses of pharmacok ineticdatawere conducted at Pfizer Inc. The
pharmacokineti cparameters were determined from pooled animal data using non-
compartmental anal ysis in Watson LIMS7.5. For PK calc ulations, 0 µg/ml was used for C0
for bothALC-0315 and AL C-0159. F or pharmacokine tic definitions and c alculations,see
Appendix 8.1. BLQ concentrations were not used in the PK calculations.
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Page 4of 154.RESULTS AND DIS CUSSION
Mean pharm acokinetic paramete rsof ALC-0315 and ALC-0159following adminis tration of
a single IVdose ofa luciferase -encoding modRNA formulated in a nLNP with an identical
lipid composition as PF -07302048 to male Wistar Han rats at 1 mg/kg are sho wn in
Supportive Ta ble6.1. Meanconcentrati on-time dataof ALC-0315 and ALC-0159from
plasma,liverhomogenates, ur ine, and feces are shown in Supportive Tables 6.2, 6.3, 6.5, and
6.6. The ratio of plasma concentr ations of AL C-0315:ALC-0159 is sh own in Supportive
Table6.4. The concentration-time profilesof plasma, liver, and fecesfollowing IV
administration of the LNP are shown in Supportive Figures 7.1,7.2,and 7.3, respectivel y.
Following a single IVdose of a lucifer ase-encoding modRNA formulated in anLNP with an
identicallipid composi tion as PF -07302048 at 1mg/kg to Wistar Han ra ts, plasma
concentrations of ALC-0315and ALC-0159 decreased rapi dly, with initial t½values of 1.62
and 1.72 h , respectivel y. ALC-0315and ALC-0159were then cleared from plasma, resulting
in terminal elimination t½of 139and 72.7 h, respectively .
Theestimated percent of dose distribute d to the liverwas~60% for AL C-0315 and ~ 20% for
ALC-0159. The percentof dose excre ted unchanged infeceswas ~1% for A LC-0315 and
~50%for ALC-0159. The percentof dose excreted unchanged in the urine was not
calculated due to values being BLQ.
5.ARCHIVING
Data presente d in this report can be found in the following locations:
ExperimentalData
E-Workbook /Root/PDM/ VBN#00701419/VR_LNP /20200801
PF-07302048_06Ju l20_072424_PLM
/Root/PDM/ VBN#00701419/VR_LNP/2020080 5
PF-07302048_06Jul20_ 072424_l iver
/Root/PDM/ VBN#00701419/VR_LNP/2 0200806
PF-07302028_06Jul20_072424_U rine
/Root/PDM/ VBN#0070 1419/VR_LNP/2020080 9
PF-07302048_06Jul20_072424_Feces
Watson LIMS PROJECT ID: PF-07302048
STUDY I D:Covidvac 072414P K
Bioanalytical Data
E-Workbook /Root/PDM/ VBN#00 701419/VR_LNP/Assay
Development/LC-MSMS method for COVID -19 Excipients
OpenLAB LAJ_PDM \COMPOUND \PF-07302048 \Covidvac_072424PK \PLM.wiff
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(b) (6)
(b) (6)
(b) (6)
(b) (6)
(b) (6)
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Page 5of 156. SUPPORTIVE TABLES
6.1.Summary of Mean PlasmaPharmacokineticParameters of ALC-0315 and ALC-0159 in MaleWistar Han Rats
Following Single IV Administration of Luciferase -encoding modRNA Formulated in anLNP with an Identical L ipid
Composition as PF-07302048 at 1mg/kg
Species(Strain) Rat (Wistar Han)
Sex/Number of Anima ls Male/ 3 animals per timepointa
Feeding Condition Fasted
Method of Administ ration IV
Dose modRNA(mg/kg) 1
Dose ALC -0159(mg/kg) 1.96
Dose ALC -0315(mg/kg) 15.3
SampleMatrix Plasma
Sampling Time Points (h post dose): Predose, 0.1, 0.25 , 0.5, 1, 3,6,24,48,96, 192, 336
Analyte ALC-0315 ALC-0159
PK Param eters: MeanbMeanb
AUCinf(µg•h/mL)c1030 99.2
AUClast(µg•h/mL) 1020 98.6
Initial t½(h)d1.62 1.74
Terminalelimination t½(h)e139 72.7
Estimated fractionof dose distributed toliver (%)f59.5 20.3
Dose in Urine (%) NCgNCg
Dosein Feces(%) 1.05 47.2
a. Non-serial sampl ing, 36 animals total.
b. Only mean PK paramet ersare reported dueto non-serial sampling.
c. Calculated using the terminal log-linear phase(determined using 48, 96, 192, and 336 hfor regressioncalculatio n).
d. ln(2)/initialelimination rat e constant (determined using 1,3, and 6 h for regression calculation ).
e.ln(2)/terminal elimination rate cons tant(determined using 48, 96, 192, and 336 f or regressioncalculation).
f. Calculated as follows: highestmeanamountin the liver (µ g)/total mean dose (µg) of ALC-0315 or ALC-0159.
g. Not calculated due to BLQ da ta.
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Page 6of 156.2. Summary of Mean and Individual Plasmaand Liver Concentrations of ALC-0315
in Male Wistar Han Rats Following Single IVAdministration of Luciferase -encoding
modRNA Formulated in an LNP with an Identi cal Lipid Composition as PF -07302048
at 1 mg/kg
Animal
NumberAnimal
weight
(g)Time
(hpostdose)Plasma
(µg/mL)Liver
(µg/g)Liver
weight (g)Total amount in
liver (µg)
1 -- Predose BLQ BLQ 5.78 BLQ
2 -- Predose BLQ BLQ 5.57 BLQ
3 -- Predose BLQ BLQ 5.89 BLQ
Mean±SD -- Predose BLQ ± NA B LQ± NA -- BLQ± NA
4 221 0.1 456 28.5 5.74 163
5 225 0.1 564 30.2 5.62 170
6 220 0.1 435 30.9 5.52 171
Mean±SD -- 0.1 485±69.2 29.9±1.23 -- 168±3.93
7 222 0.25 418 74.1 5.94 440
8 227 0.25 451 90.6 5.91 536
9 218 0.25 191 29.0 5.54 161
Mean±SD -- 0.25 353± 142 64.6±31.9 -- 379± 195
10 228 0.5 356 164 6.49 1060
11 217 0.5 333 143 5.87 839
12 224 0.5 423 140 5.03 705
Mean±SD -- 0.5 371±46.8 149±13.1 -- 869± 182
13 212 1 204 228 6.04 1380
14 221 1 208 228 6.29 1430
15 238 1 172 240 6.40 1540
Mean±SD -- 1 195±19.7 232±6.93 -- 1450±80.3
16 220 3 85.3 282 6.45 1820
17 209 3 71.2 318 5.63 1790
18 210 3 83.1 282 5.62 1580
Mean±SD -- 3 79.9±7.59 294±20.8 -- 1730 ±129
19 222 6 24.2 270 6.04 1630
20 222 6 20.2 279 5.50 1530
21 222 6 24.4 256 6.56 1680
Mean±SD -- 6 22.9±2.37 268±11.6 -- 1620±74.4
22 228 24 1.13 277 7.26 2010
23 229 24 1.11 281 7.48 2100
24 231 24 0.861 285 7.52 2140
Mean±SD -- 24 1.03±0.150 281±4.00 -- 2090±68.4
25 222 48 0.253 218 7.59 1650
26 228 48 0.339 206 7.90 1630
27 223 48 0.399 166 7.39 1230
Mean±SD -- 48 0. 330 ±0.0734 197± 27.2 -- 1500±240
28 216 96 0.188 187 7.84 1470
29 224 96 0.122 114 8.81 1000
30 242 96 0.192 97.1 9.86 957
Mean±SD -- 96 0. 167 ±0.0393 133±47.8 -- 1140 ±282
31 216 192 0.0812 66.9 9.10 609
32 212 192 0.135 60.7 8.34 506
33 233 192 0.110 90.3 9.59 866
Mean±SD -- 192 0. 109 ±0.0269 72.6±15.6 -- 660± 185
34 210 336 0.0538 53.6 10.1 541
35 213 336 0.0724 95.3 8.13 774
36 219 336 0.0801 62.2 11.0 682
Mean±SD -- 336 0.0688 ± 0.0135 7 0.4±22.0 -- 666± 118
The limit of quanti tation was 0.00488 µg/mL for plasma and 0.01953 µg/ gfor liver.
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Page 7of 156.3. Summary of Mean an dIndividual Plasmaand LiverConcentrationsof ALC-0159
in Male Wistar Han Rats Following Single IVAdministration of Luciferase -encoding
modRNA Formulated in an LNP with an Identical Lipid Composition as PF -07302048
at1 mg/kg
Animal
NumberAnimal
weight
(g)Time
(h
postdose)Plasma
(µg/mL)Liver
(µg/g)Liver weight(g)Totalamount
in liver (µg)
1 -- Predose BLQ BLQ 5.78 BLQ
2 -- Predose BLQ BLQ 5.57 BLQ
3 -- Predose BLQ BLQ 5.89 BLQ
Mean± SD -- Predose BLQ± NA BLQ ± NA -- BLQ ± NA
4 221 0.1 48.8 7.07 5.74 40.6
5 225 0.1 57.9 8.03 5.62 45.1
6 220 0.1 43.9 7.65 5.52 42.3
Mean±SD -- 0.1 50.2±7.10 7.58 ±0.483 -- 42.7 ±2.32
7 222 0.25 37.9 10.8 5.94 64.1
8 227 0.25 41.9 12.5 5.91 73.9
9 218 0.25 18.6 4.48 5.54 24.8
Mean±SD -- 0.25 32.8 ±12.5 9.26 ±4.23 -- 54.3 ±26.0
10 228 0.5 33.8 15.0 6.49 97.3
11 217 0.5 28.3 14.4 5.87 84.5
12 224 0.5 29.0 16.2 5.03 81.5
Mean±SD -- 0.5 30.4±2.99 15.2 ±0.917 -- 87.8 ±8.40
13 212 1 14.1 15.2 6.04 91.8
14 221 1 18.5 13.3 6.29 83.6
15 238 1 15.2 14.3 6.40 91.5
Mean±SD -- 1 15.9±2.29 14.3 ±0.950 -- 89.0 ±4.63
16 220 3 7.67 12.6 6.45 81.3
17 209 3 5.55 12.9 5.63 72.6
18 210 3 6.64 13.6 5.62 76.4
Mean±SD -- 3 6.62±1.06 13.0 ±0.513 -- 76.8 ±4.35
19 222 6 1.94 7.74 6.04 46.8
20 222 6 1.98 7.12 5.50 39.1
21 222 6 2.50 7.80 6.56 51.2
Mean±SD -- 6 2.14±0.312 7.55±0.376 -- 45.7 ±6.09
22 228 24 0.270 2.14 7.26 15.5
23 229 24 0.251 1.58 7.48 11.8
24 231 24 0.223 1.64 7.52 12.3
Mean±SD -- 24 0.248 ±0.0236 1.79±0.307 -- 13.2 ±2.01
25 222 48 0.113 0.565 7.59 4.29
26 228 48 0.105 0.593 7.90 4.69
27 223 48 0.0842 0.546 7.39 4.03
Mean±SD -- 48 0. 101 ±0.0149 0.568±0.0236 4.34 ±0.329
28 216 96 0.0631 0.216 7.84 1.69
29 224 96 0.0385 0.138 8.81 1.22
30 242 96 0.0524 0.148 9.86 1.46
Mean±SD -- 96 0.0513 ±0.01230.167±0.0424 -- 1.46 ±0.239
31 216 192 0.0182 0.0647 9.10 0.589
32 212 192 0.0204 0.0553 8.34 0.461
33 233 192 0.0226 0.0619 9.59 0.593
Mean±SD -- 192 0. 0204 ±0.00220 0.0606±0.00483 -- 0.548±0.0750
34 210 336 0.00568 BLQ 10.1 BLQ
35 213 336 0.00619 BLQ 8.13 BLQ
36 219 336 0.00639 BLQ 11.0 BLQ
Mean±SD -- 336 0.00609 ±0.000366 BLQ± NA -- BLQ± NA
The limit of quantita tion was 0.00488µg/mL for plasma and 0.01953µg/gfor liver.
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Page 8of 156.4. Summary of the Ratio of ALC-0315:ALC-0159in Plasma of Male Wistar Han Rat s
Following Single IV Administra tion ofLuciferase -encoding modRNA Formulated in an
LNP with an Identical Lipid Composition as PF -07302048 at 1 mg/kg
Timepostdose
(h)Meanplasma
concentration
ALC-0315(µg/mL)Meanplasma
concentration
ALC-0159(µg/mL)Ratio
(ALC-0315)/(ALC -0159)a
0 BLQ BLQ NA
0.1 485 50.2 9.66
0.25 353 32.8 10.8
0.5 371 30.4 12.2
1 195 15.9 12.3
3 79.9 6.62 12.1
6 22.9 2.14 10.7
24 1.03 0.248 4.15
48 0.330 0.101 3.27
96 0.167 0.0513 3.26
192 0.109 0.0204 5.34
336 0.0688 0.00609 11.3
The limit of quantitation was 0.004 88µg/mL.
a. Ratio prior to injection is 7.8 ( 15.3 mg/kg/1.96 mg/ kg)
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Page 9of 156.5. Summary of Mean an d Individual Urine Conc entrations of ALC-0315and
ALC-0159in Male Wistar Han Rat s Following Single IVAdministration of
Luciferase -encoding mod RNA Formulated in an LNP with an Identical Lipid
Composition as PF-07302048 at1 mg/kg
ALC-0315 ALC-0159
Individual Animal MeanSD Individual Animal MeanSD
Animal
No. 34Animal
No. 35Animal
No. 36Animal
No. 34Animal
No.35Animal
No.36
Animal We ight (g) 210 213 219 -- 210 213 219 --
Time (hpostdose) Urine Concentration (ng/mL)
Predose BLQBLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
0-24 BLQ BLQ BLQBLQ±NABLQ BLQBLQBLQ±NA
24-48 BLQ BLQ BLQBLQ±NABLQBLQ BLQBLQ±NA
48-72 BLQ BLQ BLQBLQ±NABLQ BLQBLQBLQ±NA
72-96 BLQ BLQ BLQBLQ±NABLQBLQ BLQBLQ±NA
96-120 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
120-144 BLQ BLQBLQBLQ±NABLQ BLQ BLQBLQ±NA
144-168 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
168-192 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
192-216 BLQ BLQ BLQBLQ±NABLQ BLQBLQBLQ±NA
216-240 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
240-264 BLQ BLQ BLQBLQ±NABLQ BLQBLQBLQ±NA
264-288 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
288-312 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
312-336 BLQ BLQ BLQBLQ±NABLQ BLQ BLQBLQ±NA
Time (h) Urine Volume (m L)
Predose 22.0a17.0 15.0a-- 22.0a17.0 15.0a--
0-24 11.0 10.0 15.0 -- 11.0 10.0 15.0 --
24-48 9.00 7.5012.0 -- 9.00 7.5012.0 --
48-72 10.0 10.0 10.0 -- 10.0 10.0 10.0 --
72-96 12.010.0 9.00 -- 12.0 10.0 9.00 --
96-120 15.0 12.0 12.5 -- 15.0 12.0 12.5 --
120-144 14.5 11.512.5 -- 14.5 11.5 12.5 --
144-168 13.0 9.00 10.0 -- 13.0 9.00 10.0 --
168-192 15.0b12.5 14.5 -- 15.0b12.5 14.5 --
192-216 13.0 8.00 12.5 -- 13.0 8.00 12.5 --
216-240 12.0 8.00 12.0 -- 12.08.00 12.0 --
240-264 15.0 14.0 16.0 -- 15.0 14.0 16.0 --
264-288 9.50 8.00 12.0 -- 9.50 8.00 12.0 --
288-312 13.0 10.0 15.0 -- 13.010.0 15.0 --
312-336 16.0 10.0 15.0 -- 16.0 10.0 15.0 --
Thelimit of quantitation w as 0.00488 µg/mL
a. Possible wa ter contamination.
b. Urine over flowed.
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Page 10of 156.6.Summary of Mean an dIndividual Fecal Concentrations of ALC-0315 and
ALC-0159in Male Wist ar Han Rats Following Singl e IVAdministration of
Luciferase -encoding mod RNA Formulated in an LNP with an Identical Lipid
Composition as PF-07302048 at 1 mg/kg
ALC-0315 ALC-0159
Individual Animal MeanSD Individual Animal MeanSD
No. 34No. 35No. 36 No. 34No.35No.36
Animal Weight
(g)210213219 -- 210 213 219 --
Time (hpostdose) FecalConcentration ( µg/g)
Predose BLQBLQ BLQ BLQ ± NA BLQ BLQ BLQ BLQ± NA
0-24 1.500.474 0.397 0.790±0.616 38.928.015.2 27.4±11.9
24-48 0.9720.432 0.778 0.727±0.274 2.17 5.64 4.79 4.20±1.81
48-72 0.355 0.572 1.33 0.752±0.512 0.7100.952 1.330.997±0.312
72-96 0.1670.2940.450 0.304±0.142 0.401 0.389 0.421 0.404±0.0162
96-120 0.369 0.114 0.173 0.219±0.133 0.332 0.269 0.217 0.273±0.0576
120-144 0.239 0.0881 0.113 0.147±0.08090.375 0.171 0.143 0.230±0.127
144-168 0.08880.1000.175 0.121±0.0469 0.3000.1570.228 0.228±0.0715
168-192 0.110 0.07830.1290.106±0.0256 0.145 0.124 0.1470.139±0.0127
192-216 0.07900.0809 0.1830.114±0.0595 0.0904 0.163 0.1460.133±0.0380
216-240 0.142 0.1520.101 0.132±0.0270 0.155 0.182 0.116 0.151±0.0332
240-264 0.07810.0764 0.1350.0965±0.0334 0.1090.07830.142 0.110±0.0319
264-288 0.0947 0.06350.122 0.0934±0.0293 0.07540.0641 0.109 0.0828±0.0234
288-312 0.0457 0.0758 0.107 0.0762±0.0307 0.106 0.05800.0982 0.0874±0.0258
312-336 0.02900.06410.09930.0641±0.03520.0605 0.0494 0.08540.0651±0.0184
Time (h) Feces weight(g)
Predose 4.80 4.40 6.40 -- 4.80 4.40 6.40 --
0-24 2.50 7.90 5.50 -- 2.50 7.90 5.50 --
24-48 8.90 5.508.70 -- 8.90 5.50 8.70 --
48-72 11.5 8.60 13.1 -- 11.5 8.60 13.1 --
72-96 11.8 13.110.5 -- 11.8 13.110.5 --
96-120 13.9 10.6 13.2 -- 13.9 10.6 13.2 --
120-144 10.9 12.9 12.6 -- 10.9 12.9 12.6 --
144-168 13.5 15.3 8.20 -- 13.5 15.3 8.20 --
168-192 8.40 15.4 10.5 -- 8.40 15.4 10.5 --
192-216 13.0 11.38.00 -- 13.0 11.3 8.00 --
216-240 10.1 11.1 9.80 -- 10.1 11.1 9.80 --
240-264 10.4 11.4 9.40 -- 10.4 11.4 9.40 --
264-288 11.1 11.2 8.40 -- 11.1 11.2 8.40 --
288-312 11.7 11.69.70 -- 11.7 11.6 9.70 --
312-336 12.0 11.5 7.20 -- 12.0 11.5 7.20 --
Time (h) Amount ex creted in feces (µg)
Predose BLQ BLQBLQ BLQ ± NA BLQ BLQ BLQ BLQ± NA
0-24 3.75 3.74 2.18 3.23±0.903 97.3 221 83.6 134±75.8
24-48 8.65 2.38 6.77 5.93±3.22 19.3 31.0 41.7 30.7±11.2
48-72 4.084.9217.4 8.81±7.47 8.17 8.19 17.411.3±5.34
72-96 1.97 3.85 4.73 3.52± 1.41 4.735.10 4.42 4.75±0.338
96-120 5.13 1.21 2.28 2.87±2.03 4.61 2.85 2.86 3.44±1.01
120-144 2.61 1.14 1.42 1.72±0.778 4.09 2.21 1.80 2.70±1.22
144-168 1.20 1.531.441.39±0.171 4.05 2.40 1.87 2.77±1.14
168-192 0.924 1.21 1.351.16±0.2191.221.91 1.54 1.56 ±0.346
192-216 1.030.914 1.46 1.14±0.290 1.18 1.84 1.17 1.40±0.387
216-240 1.43 1.690.990 1.37±0.353 1.57 2.02 1.14 1.57±0.442
240-264 0.8120.8711.270.984±0.249 1.13 0.8931.33 1.12 ±0.221
264-288 1.050.711 1.02 0.929±0.189 0.837 0.718 0.916 0.823±0.100
288-312 0.535 0.879 1.04 0.817±0.257 1.24 0.673 0.953 0.955±0.284
312-336 0.348 0.737 0.715 0.600±0.2190.726 0.568 0.6150.636±0.0811
Thelimitof quantitationwas0.006592 µg/mL.
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Page 11of 157.SUPPORTIVE FIGURES
7.1.Mean PlasmaConcentrations of ALC-0315 and ALC -0159in Male Wistar Han
RatsFollowing Single IVAdministrat ionof Luciferase -encoding mod RNA Formulated
in anLNP with an Identical Lipid Composition as PF -07302048at 1 mg/kg
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Page 12of 157.2.Mean Liver Concentrations o f ALC-0315 and ALC-0159in Male Wistar Han Rats
Following Single IVAdministration of Luciferase -encoding mod RNA Formulated in an
LNP with an Identical Lipid Composition as PF-07302048 at 1 mg/kg
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Page 13of 157.3.Mean FecalConcentrations of ALC-0315 and ALC -0159in MaleWistar Han Rats
Following Single IV Administration of Luciferase-encoding mod RNA Formulated in an
LNP with an Identical Lipid Composition as PF-07302048 at1 mg/kg
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Page 14of 158.SUPPORTIVE APPENDIX
8.1.Abbreviationsand Pharmacokinetic Calculations
-- Data not available
AUCinf Area under the plasmadrug concentration -time curve from 0to infinite time. AUCtplus
extrapolated a readetermined bydividing plasmaconcentrationat tby the slopeof the
terminal log-linear phase.
AUClast Area under the plasmadrug concentration-time curve from 0 to the last quanti fiable tim e
point. Determined using the linear t rapezoidal method.
BLQ Below the limitof quantitation
K2EDTA Potassium ethyl ene diamine t etraacetic acid
ID Identificati on
Initial t½ Half-life.
ln(2)/initialelimination rate constant
ISTD Internal standard
IV Intravenous
LNP Lipid nanoparticle
LC-MS/MS Liquid chrom atography -tandem mass spectro metry
MRM Multiple reaction monitoring
NA Notapplicable
NC Not calcula ted
PEG Polyethylene glycol
PK Pharmacokinetic
SD Standard dev iation
Terminal
eliminationt½Half-life.
ln(2)/terminal elimination rate constant
Dose in Feces(%)Fecalexcretion.
(Mean µg of analyte in feces/ Mean µgof analyte administered) × 100
Dose in Urine(%)Urinary excretion.
(Mean µg of analytein urine/ Mean µg of analyte administered) × 100
9. OTHERCONTRIBUTING SCIENTISTS
The following s cientists were involved in the conduct of the PK portion o f thisstudyand are
responsibleforthe scientific content of thi s research report.
ADME Associate/
Portfolio Support Group
Scientist
Bioanalytical Associate
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Page 15of 1510. CHANGE HISTORY
Version Sections Revisions
1.0 NA New document
2.0 Summary , Section 3.1,
Results and Discussion,
Supportive Table 6.1Updated the dosesof ALC-0159 and
ALC-0315 and the percent of dose distribute d
to the liverand excreted i n feces.
11.APPROVAL SIG NATURE S
Theauthor and approver are responsibleforproviding a true representation of the d ata.
Report Author
Pharmacoki netics, Dynamics an d Metabolism, Pfizer, Groton,CT, USA
,
Report Approver
Pharmacokinet ics, Dynamics and Metabolism,Pfizer, San Diego, CA, USA
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Document Approval Record
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