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BNT162b2
2.6.5 Pharmacokinetics Tabulated Summary
CONFIDENTIAL
Page 1MODULE 2.6.5. PHARMA COKINETICS TABULATED SUMMARY
This document contains confidential information belonging to BioNTech /Pfizer. Except as may be
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2.6.5 Pharmacokinetics Tabulated Summary
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Page 22.6.5.1. PHARMACOKINETICS OVE RVIEW Test Article: BNT162b2
Type of Study Test System Test item Method of
AdministrationTesting Facility Report Number
Single Dose Pharmacokinetics
Single Dose
Pharmacokinetics and
Excretion in Urine and Feces
of ALC-0159 and ALC -0315Rat (Wistar Han) modRNA encoding
luciferase
formulated in LNP
comparable to
BNT162b2IV bolus Pfizer IncaPF-07302048_06Jul20_072424
Distribution
In Vivo Distribution Mice (BALB/c)modRNA encoding
luciferase
formulated in LNP
comparable to
BNT162b2IM Injection BioNTechbR-20-0072
In Vivo Distribution Rat (Wistar Han) modRNA encoding
luciferase
formulated in LNP
comparable to
BNT162b2 w ith
trace amounts of
[3H]-CHE as
non-diffusible labelIM Injection 185350
Metabolism
In Vitro and In Vivo Metabolism
In Vitro Metabolic Stability
of ALC-0315in Liver
MicrosomesMouse (CD-1/ICR), rat
(Sprague Dawley and
Wistar Han ), monkey
(Cynomolgus ), and
human liver microsomesALC-0315 In vitro 01049-20008
In Vitro Metabolic Stability
of ALC-0315in Liver S9Mouse (CD-1/ICR), rat
(Sprague Dawley ),
monkey (Cynomolgus ),
and human S9 liver
fractionsALC-0315 In vitro 01049-20009
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(b) (4)
(b) (4)
(b) (4)
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Page 32.6.5.1. PHARMACOKINETICS OVE RVIEW Test Article: BNT162b2
Type of Study Test System Test item Method of
AdministrationTesting Facility Report Number
In Vitro Metabolic Stability
of ALC-0315in HepatocytesMouse (CD -1/ICR), rat
(Sprague Dawley and
Wistar Han), monkey
(Cynomolgus), and
human hepatocytesALC-0315 In vitro 01049-20010
In Vitro Metabolic Stability
of ALC-0159 in Liver
MicrosomesMouse (CD -1/ICR), rat
(Sprague Dawley and
Wistar Han), monkey
(Cynomolgus), and
human liver microsomesALC-0159 In vitro 01049-20020
In Vitro Metabolic Stability
of ALC-0159 in Liver S9Mouse (CD -1/ICR), rat
(Sprague Dawley),
monkey (Cynomolgus),
and human S9 fractionsALC-0159 In vitro 01049-20021
In Vitro Metabolic Stability
of ALC-0159 in HepatocytesMouse (CD -1/ICR), rat
(Sprague Dawley and
Wistar Han), monkey
(Cynomolgus), and
human hepatocytesALC-0159 In vitro 01049-20022
Biotransformation of
ALC-0159 and ALC -0315 In
Vitro and In Vivo in RatsIn vitro:
CD-1mouse,Wistar
Hanrat,cynomolgus
monkey, and human
blood, liver S9 fractions
and hepatocytes
In vivo: male Wistar Han
ratsALC-0315 and
ALC-0159In vitro or
IV (in vivo in
rats)Pfizer IncePF-07302048_05Aug20_043725
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(b) (4)
(b) (4)
(b) (4)
(b) (4)
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Page 42.6.5.1. PHARMACOKINETICS OVE RVIEW Test Article: BNT162b2
Type of Study Test System Test item Method of
AdministrationTesting Facility Report Number
ALC-0159 = 2-[(polyethylene glycol) -2000]-N,N-ditetradecylacetamide), a proprietary polyethylene glycol-lipidincluded as an excipient in the LNP formulation
used in BNT162b2 ; ALC-0315 = (4-hydroxybutyl)azanediyl)bis(hexane -6,1-diyl)bis(2-hexyldecanoate), a proprietary aminolipid included as an excipient in the
LNP formulation used in BNT162b2 ; [3H]-CHE= radiolabeled [ cholesteryl -1,2-3H(N)]-cholesteryl hexadecyl ether;IM = Intramuscular; IV = Intravenous; LNP
= lipid nanoparticles ; S9 = Supernatant fraction obtained from liver homogenate by centrifuging at 9000 g .
a. La Jolla, California.
b. Mainz, Germany.
e. Groton, Connecticut.
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Page 52.6.5.3. PHARMACOKIN ETICS:
PHARMACOKINETICS AFTER A SINGLE DOSETest Article: modRNA encoding luciferase in LNP
Report Number: PF-07302048_06Jul20_072424
Species (Strain) Rat (Wistar Han)
Sex/Number of Animals Male/ 3 animals per timepointa
Feeding Condition Fasted
Method of Administration IV
Dose modRNA (mg/kg) 1
Dose ALC -0159 (mg/kg) 1.96
Dose ALC -0315 (mg/kg) 15.3
Sample Matrix Plasma, liver, urine and feces
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 Parameters: MeanbMeanb
AUCinf(µg•h/mL)c1030 99.2
AUClast(µg•h/mL) 1020 98.6
Initial t ½(h)d1.62 1.74
Terminal elimination t ½(h)e139 72.7
Estimated fraction of dose distributed to liver (%)f59.5 20.3
Dose in Urine (%) NCgNCg
Dose in Feces (%)h1.05 47.2
ALC-0159 = 2-[(polyethylene glycol) -2000]-N,N-ditetradecylacetamide ), a proprietary polyethylene glycol -lipid included as an excipient in the LNP formulation
used in BNT162b2 ; ALC-0315 = (4 -hydroxybutyl)azanediyl)bis(hexane -6,1-diyl)bis(2-hexyldecanoate), a proprietary aminolipid included as an excipient in the
LNP formul ation used in BNT162b2 ; AUCinf = Area under the plasma drug concentration -time curve from 0 to infinite time; AUClast= Area under the plasma
drug concentration -time curve from 0 to the last quantifiable time point; BLQ = Below the limit of quantitation; LNP = Lipid nanoparticle;
modRNA=Nucleoside modified messenger RNA ; PK = Pharmacokinetics; t ½ = Half-life.
a. Non-serial sampling, 36 animals total.
b. Only mean PK parameters are reported due to non -serial sampling.
c. Calculated using the terminal log-linear phase (determined using 48, 96, 192, and 336 h for regression calculation).
d. ln(2)/initial elimination rate constant (determined using 1, 3, and 6 h for regression calculation).
e. ln(2)/terminal elimination rate constant (determined using 48, 96, 192, and 336 hfor regression calculation).
f. Calculated as follows: highest mean amount in the liver ( µg)/total mean dose ( µg) of ALC -0315 or ALC -0159.
g. Not calculated due to BLQ data.
h. Fecal excretion, calculated as: (mean µg of analyte in feces/ mean µg of analyte administered) × 100
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Page 62.6.5.5A. PHARMACOKINETICS: ORGAN
DISTRIBUTION Test Article: modRNA encoding luciferase in LNP
Report Number: R-20-0072
Species (Strain): Mice (BALB/c)
Sex/Number of Animals: Female/3 per group
Feeding Condition: Fed ad libitum
Vehicle/Formulation: Phosphate-buffered saline
Method of Administration: Intramuscular injection
Dose (mg/kg): 1 µg/hind leg in gastrocnemius muscle (2 µg total)
Number of Doses: 1
Detection: Bioluminescence measurement
Sampling Time (hour): 6, 24, 48, 7 2hours; 6 and 9 days post -injection
Time point Total Mean Biolum inescence signal (photons/second) Mean Biolum inescence signal in
the liver (photons/second)
Buffer control modRNA Luciferase in LNP modRNA Luciferase in LNP
6 hours 1.28×1051.26×1094.94×107
24 hours 2.28×1057.31×1082.4×106
48 hours 1.40×1052.10×108Below detectiona
72 hours 1.33×1057.87×107Below detectiona
6 days 1.62×1052.92×106Below detectiona
9 days 7.66×1045.09×105Below detectiona
LNP = Lipid nanoparticle; modRNA =Nucleoside modified messenger RNA.
a. At or below the background level of the buffer control.
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Page 72.6.5.5B. PHARMACOK INETICS: ORGAN
DISTRIBUTION CONTINUEDTest Article: [3H]-Labelled LNP -mRNAformulation c ontaining
ALC-0315 and ALC -0159
Report Number: 185350
Species (Strain): Rat (Wistar Han)
Sex/Number of Animals: Male and f emale/3 animals/sex/timepoint (21 animals/sex total for the 50 µg dose)
Feeding Condition: Fed ad libitum
Method of Administration: Intramuscular injection
Dose: 50µg[3H]-08-A01-C0(lot # NC -0552-1)
Number of Doses: 1
Detection: Radioactivity quantitation using liquid scintillation counting
Sampling Time (hour): 0.25, 1, 2, 4, 8, 24, and 48 hourspost-injection
Sample Mean total lipid concentration (µg lipid equivalent/g (or mL)
(males and fem ales combined)% of adm inistered dose (males and fem ales combined)
0.25 min 1 h 2 h 4 h 8 h 24 h 48 h 0.25 min 1 h 2 h 4 h 8 h 24 h 48 h
Adiposetissue 0.057 0.100 0.126 0.128 0.093 0.084 0.181 -- -- -- -- -- -- --
Adrenalglands 0.271 1.48 2.72 2.89 6.80 13.8 18.2 0.001 0.007 0.010 0.015 0.035 0.066 0.106
Bladder 0.041 0.130 0.146 0.167 0.148 0.247 0.365 0.000 0.001 0.001 0.001 0.001 0.002 0.002
Bone (femur) 0.091 0.195 0.266 0.276 0.340 0.342 0.687 -- -- -- -- -- -- --
Bone marrow
(femur)0.479 0.960 1.24 1.24 1.84 2.49 3.77 -- -- -- -- -- -- --
Brain 0.045 0.100 0.138 0.115 0.073 0.069 0.068 0.007 0.013 0.020 0.016 0.011 0.010 0.009
Eyes 0.010 0.035 0.052 0.067 0.059 0.091 0.112 0.000 0.001 0.001 0.002 0.002 0.002 0.003
Heart 0.282 1.03 1.40 0.987 0.790 0.451 0.546 0.018 0.056 0.084 0.060 0.042 0.027 0.030
Injection site 128 394 311 338 213 195 165 19.9 52.6 31.6 28.4 21.9 29.1 24.6
Kidneys 0.391 1.16 2.05 0.924 0.590 0.426 0.425 0.050 0.124 0.211 0.109 0.075 0.054 0.057
Large intestine 0.013 0.048 0.093 0.287 0.649 1.10 1.34 0.008 0.025 0.065 0.192 0.405 0.692 0.762
Liver 0.737 4.63 11.0 16.5 26.5 19.2 24.3 0.602 2.87 7.33 11.9 18.1 15.4 16.2
Lung 0.492 1.21 1.83 1.50 1.15 1.04 1.09 0.052 0.101 0.178 0.169 0.122 0.101 0.101
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Page 82.6.5.5B. PHARMACOK INETICS: ORGAN
DISTRIBUTION CONTINUEDTest Article: [3H]-Labelled LNP -mRNAformulation c ontaining
ALC-0315 and ALC -0159
Report Number: 185350
Sample Total Lipid concentration (µg lipid equivalent/g [or mL])
(males and fem ales combined)% of Administered Dose (males and fem ales combined)
0.25 min 1 h 2 h 4 h 8 h 24 h 48 h 0.25 min 1 h 2 h 4 h 8 h 24 h 48 h
Lymph node
(mandibular)0.064 0.189 0.290 0.408 0.534 0.554 0.727 -- -- -- -- -- -- --
Lymph node
(mesenteric)0.050 0.146 0.530 0.489 0.689 0.985 1.37 -- -- -- -- -- -- --
Muscle 0.021 0.061 0.084 0.103 0.096 0.095 0.192 -- -- -- -- -- -- --
Ovaries
(females)0.104 1.34 1.64 2.34 3.09 5.24 12.3 0.001 0.009 0.008 0.016 0.025 0.037 0.095
Pancreas 0.081 0.207 0.414 0.380 0.294 0.358 0.599 0.003 0.007 0.014 0.015 0.015 0.011 0.019
Pituitary gland 0.339 0.645 0.868 0.854 0.405 0.478 0.694 0.000 0.001 0.001 0.001 0.000 0.000 0.001
Prostate
(males)0.061 0.091 0.128 0.157 0.150 0.183 0.170 0.001 0.001 0.002 0.003 0.003 0.004 0.003
Salivary
glands0.084 0.193 0.255 0.220 0.135 0.170 0.264 0.003 0.007 0.008 0.008 0.005 0.006 0.009
Skin 0.013 0.208 0.159 0.145 0.119 0.157 0.253 -- -- -- -- -- -- --
Small intestine 0.030 0.221 0.476 0.879 1.28 1.30 1.47 0.024 0.130 0.319 0.543 0.776 0.906 0.835
Spinal cord 0.043 0.097 0.169 0.250 0.106 0.085 0.112 0.001 0.002 0.002 0.003 0.001 0.001 0.001
Spleen 0.334 2.47 7.73 10.3 22.1 20.1 23.4 0.013 0.093 0.325 0.385 0.982 0.821 1.03
Stomach 0.017 0.065 0.115 0.144 0.268 0.152 0.215 0.006 0.019 0.034 0.030 0.040 0.037 0.039
Testes (males) 0.031 0.042 0.079 0.129 0.146 0.304 0.320 0.007 0.010 0.017 0.030 0.034 0.074 0.074
Thymus 0.088 0.243 0.340 0.335 0.196 0.207 0.331 0.004 0.007 0.010 0.012 0.008 0.007 0.008
Thyroid 0.155 0.536 0.842 0.851 0.544 0.578 1.00 0.000 0.001 0.001 0.001 0.001 0.001 0.001
Uterus
(females)0.043 0.203 0.305 0.140 0.287 0.289 0.456 0.002 0.011 0.015 0.008 0.016 0.018 0.022
Whole blood 1.97 4.375.40 3.051.310.909 0.420 -- -- -- -- -- -- --
Plasma 3.978.13 8.90 6.50 2.36 1.78 0.805 -- -- -- -- -- -- --
Blood:Plasma
ratioa0.815 0.515 0.550 0.510 0.555 0.530 0.540 -- -- -- -- -- -- --
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Page 92.6.5.5B. PHARMACOK INETICS: ORGAN
DISTRIBUTION CONTINUEDTest Article: [3H]-Labelled LNP -mRNAformulation c ontaining
ALC-0315 and ALC -0159
Report Number: 185350
--= Not applicable, partial tissue taken; [3H]-08-A01-C0 = An aqueous dispersion of LNP s, including ALC -0315, ALC -0159,distearoylphosphatidylcholine,
cholesterol , mRNAencoding luciferase andtrace amounts of radiolabeled [Cholesteryl -1,2-3H(N)]-Cholesteryl Hexadecyl Ether, a nonexchangeable, non-
metabolizable lipid marker used to monitor the disposition of the LNPs; ALC-0159 = 2-[(polyethylene glycol) -2000]-N,N--ditetradecylacetamide), a prop rietary
polyethylene glycol -lipid included as an excipient in the LNP formulation used in BNT162b2 ; ALC-0315=(4--hydroxybutyl)azanediyl)bis(hexane -6,1-
diyl)bis(2 -hexyldecanoate), a proprietary aminolipid included as an excipient in the LNP formulation us ed in BNT162b2 ; LNP = Lipid nanoparticle;
mRNA= messenger RNA.
a. The mean male and female blood:plasma values were first calculated separately and this value represents the mean of the two values .
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Page 102.6.5.9. PHARMACOKI NETICS: METABOLISM I N VIVO,
RATTest Article: modRNA encoding luciferase in LNP
Report Number: PF-07302048_05Aug20_043725
Species (Strain): Rat (Wistar Han)
Sex/ Number of animals Male/ 36 animals total for plasma and liver, 3 animals for urine and feces
Method of Administration: Intravenous
Dose (mg/kg): 1
Test System: Plasma, Urine, Feces , Liver
Analysis Method: Ultrahigh performance liquid chromatography /mass spectrometry
Biotransform ation m/z Metabolites of ALC- 0315 Detected
Plasma Urine Feces Liver
N-dealkylation, oxidation 102.0561aND ND ND ND
N-Dealkylation, oxidation 104.0706bND ND ND ND
N-dealkylation, oxidation 130.0874aND ND ND ND
N-Dealkylation, oxidation 132.1019bND ND ND ND
N-dealkylation, hydrolysis, oxidation 145.0506aND ND ND ND
Hydrolysis (acid) 255.2330a+ ND ND ND
Hydrolysis, hydroxylation 271.2279aND ND ND ND
Bis-hydrolysis (amine) 290.2690b+ + + +
Hydrolysis, glucuronidation 431.2650aND ND ND ND
Bis-hydrolysis (amine), glucuronidation 464.2865aND ND ND ND
Bis-hydrolysis (amine), glucuronidation 466.3011bND + ND ND
Hydrolysis (amine) 528.4986b+ ND ND +
Hydrolysis (amine), Glucuronidation 704.5307bND ND ND ND
Oxidation to acid 778.6930aND ND ND ND
Oxidation to acid 780.7076bND ND ND ND
Hydroxylation 782.7232bND ND ND ND
Sulfation 844.6706aND ND ND ND
Sulfation 846.6851bND ND ND ND
Glucuronidation 940.7458aND ND ND ND
Glucuronidation 942.7604bND ND ND ND
Note: Both theoretical and observed metabolites are included.
m/z = mass to charge ratio; ND = Not detected; + = minor metabolite as assessed by ultraviolet detection .
a. Negative ion mode.
b. Positive ion mode .
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Page 112.6.5.10A. PHARMACOK INETICS: METABOLISM IN VITROTest Article: ALC -0315
Report Number s: 01049-20008
01049-20009
01049-20010
Type of Study: Stability of ALC-0315 InVitro
Study System: Liver Microsomes + NADPH S9 Fraction + NADPH, UDPGA, and
alamethicinHepatocytes
ALC-0315
Concentration:1µM 1µM 1µM
Duration of
Incubation ( min):120 min 120 min 240 min
Analysis Method: Ultra-high performance liquid chromatography -tandem mass spectrometry
Incubation time
(min)Percent ALC -0315 remaining
Liver Microsomes Liver S9 Fraction Hepatocytes
Mouse
(CD-1/ICR)Rat
(SD)Rat
(WH)Monkey
(Cyno)Human Mouse
(CD-1/ICR)Rat
(SD)Monkey
(Cyno)Human Mouse
(CD-1/ICR)Rat
(SD)Rat
(WH)Monkey
(Cyno)Human
0 100.00100.00100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00100.00 100.00 100.00
15 98.77 94.39 96.34 97.96 100.24 97.69 98.85 99.57 95.99 -- -- -- -- --
30 97.78 96.26 97.32 96.18 99.76 97.22 99.62 96.96 97.32 101.15 97.75102.70 96.36100.72
60 100.49 99.73 98.54 100.00 101.45 98.61 99.62 99.13 94.98 100.77 98.50102.32 97.82 101.44
90 97.78 98.66 94.15 97.96 100.48 98.15 98.85 98.70 98.33 101.92 99.25103.09 100.0 100.36
120 96.54 95.9993.66 97.71 98.31 96.76 98.46 99.57 99.33 98.85 97.38 99.61 96.36 100.72
180 -- -- -- -- -- -- -- -- -- 101.15 98.88103.47 95.64 98.92
240 -- -- -- -- -- -- -- -- -- 99.62 101.12 100.00 93.82 99.64
t½ (min) >120 >120 >120 >120 >120 >120 >120 >120 >120 >240 >240>240>240 >240
-- = Data not available; ALC-0315 = (4 -hydroxybutyl)azanediyl)bis(hexane-6,1 -diyl)bis(2 -hexyldecanoate), a proprietary aminolipid included as an excipient in the lipid
nanoparticle formulation used in BNT162b2 ;Cyno=Cynomolgus; NADPH=Reduced form of nicotinamide adenine dinucleotide phosphate; NC=not calculated ; SD = Sprague
Dawley; t½ = half-life; WH = Wistar -Han; UDPGA= uridine-diphosphate -glucuronic acid trisodium salt .
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Page 122.6.5.10B.PHARMACOKINETICS: METABOLISM IN VITRO CONTINUEDTest Article: ALC -0159
Report Numbers: 01049-20020
01049-20021
01049-20022
Type of Study: Stability of ALC-0159InVitro
Study System: Liver Microsomes + NADPH S9 Fraction + NADPH, UDPGA, and
alamethicinHepatocytes
ALC-0159
Concentration:1µM 1µM 1µM
Duration of
Incubation ( min):120 min 120 min 240 min
Analysis Method: Ultra-high performance liquid chromatography -tandem mass spectrometry
Incubation time
(min)Percent ALC -0159remaining
Liver Microsomes Liver S9 Fraction Hepatocytes
Mouse
(CD-1/ICR)Rat
(SD)Rat
(WH)Monkey
(Cyno)Human Mouse
(CD-1/ICR)Rat
(SD)Monkey
(Cyno)Human Mouse
(CD-1/ICR)Rat
(SD)Rat
(WH)Monkey
(Cyno)Human
0 100.00100.00100.00100.00 100.00 100.00 100.00100.00 100.00 100.00 100.00100.00100.00100.00
15 82.27 101.24112.11100.83 99.59 98.93 84.38 91.30 106.73 -- ---- -- --
30 86.40 93.78102.69 85.12 92.28 91.10 90.87 97.96 107.60 100.85 93.37113.04 90.23 106.34
60 85.54 98.34105.38 86.36 95.53 102.85 97.97 105.56 104.97 94.92 91.81105.07 92.93 101.58
90 85.41 95.44100.90 94.63 97.97 90.75 93.51 108.33 109.36 94.28 90.25112.80 94.59 92.67
120 95.87 97.10108.97 93.39 93.09 106.76 92.70 105.74 119.59 87.08 89.47104.11 97.51 96.04
180 -- -- -- -- -- -- -- -- -- 94.92 93.96102.90 89.81 93.66
240 -- -- -- -- -- -- -- -- -- 102.75 94.9398.79 92.93 102.57
t½ (min) >120 >120 >120 >120 >120 >120 >120 >120 >120 >240 >240>240>240>240
-- = Data not available; ALC-0159 = 2 -[(polyethylene glycol) -2000]-N,N -ditetradecylacetamide), a proprietary polyethylene glycol -lipid included as an excipient in the lipid
nanoparticle formulation used in BNT162b2 ; Cyno=Cynomolgus; NADPH=Reduced form of nicotinamide adenine dinucleotide phosphate; N C =notcalculated ; SD = Sprague
Dawley; WH = Wistar -Han; UDPGA= uridine-diphosphate -glucuronic acid trisodium salt .
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Page 132.6.5.10C. PHARMACOKINETICS: METABOLISM
IN VITROCONTINUEDTest Article: ALC-0315
Report Number: PF-07302048_05Aug20_043725
Type of study Metabolism of ALC -0315 InVitro
Study system Blood Hepatocytes Liver S9 Fraction
ALC-0315 concentration 10 µM 10 µM 10 µM
Duration of incubation 24 h 4h 24 h
Analysis Method: Ultrahigh performance liquid chromatography/ mass spectrometry
Biotransformation m/z Blood Hepatocytes Liver S9 Fraction
Mouse RatMonkeyHuman Mouse RatMonkeyHuman Mouse RatMonkeyHuman
N-dealkylation, oxidation 102.0561aNDNDND ND ND ND ND ND NDND ND ND
N-Dealkylation, oxidation 104.0706bNDNDND ND ND ND ND ND NDND ND ND
N-dealkylation, oxidation 130.0874aNDNDND ND ND ND ND ND NDND ND ND
N-Dealkylation, oxidation 132.1019bNDNDND ND ND ND ND ND NDND ND ND
N-dealkylation, hydrolysis, oxidation 145.0506aNDNDND ND ND ND ND ND NDND ND ND
Hydrolysis (acid) 255.2330a++ND ND + + + + ++ND +
Hydrolysis, hydroxylation 271.2279aNDNDND ND ND ND ND ND NDND ND ND
Bis-hydrolysis (amine) 290.2690b++ND ND ND ND ND ND NDND +ND
Hydrolysis, glucuronidation 431.2650aNDNDND ND ND ND ND ND NDND ND ND
Bis-hydrolysis (amine), glucuronidation 464.2865aNDNDND ND ND ND ND ND NDND ND ND
Bis-hydrolysis (amine), glucuronidation 466.3011bNDNDND ND ND ND ND ND NDND ND ND
Hydrolysis (amine) 528.4986bND +ND ND ND ND ND ND NDND +ND
Hydrolysis (amine), glucuronidation 704.5307bNDNDND ND ND ND ND ND NDND ND ND
Oxidation to acid 778.6930aNDNDND ND ND ND ND ND NDND ND ND
Oxidation to acid 780.7076bNDNDND ND ND ND ND ND NDND ND ND
Hydroxylation 782.7232bNDNDND ND ND ND ND ND NDND ND ND
Sulfation 844.6706aNDNDND ND ND ND ND ND NDND ND ND
Sulfation 846.6851bNDNDND ND ND ND ND ND NDND ND ND
Glucuronidation 940.7458aNDNDND ND ND ND ND ND NDND ND ND
Glucuronidation 942.7604bNDNDND ND ND ND ND ND NDND ND ND
Note: Both theoretical and observed metabolites are included.
m/z = mass to charge ratio; ND = Not detected; + = metabolite present.
a. Negative ion mode .
b. Positive ion mode .
090177e196137069\Approved\Approved On: 21-Jan-2021 23:22 (GMT)
FDA-CBER-2021-5683-0013919
BNT162b2
2.6.5 Pharmacokinetics Tabulated Summary
CONFIDENTIAL
Page 142.6.5.10D. PHARMACOKINETICS: METABOLISM
IN VITROCONTINUEDTest Article: ALC-0159
Report Number: PF-07302048_05Aug20_043725
Type of study Metabolism of ALC -0159InVitro
Study system Blood Hepatocytes Liver S9 Fraction
ALC-0159concentration 10 µM 10 µM 10 µM
Duration of incubation 24 h 4h 24 h
Analysis Method: Ultrahigh performance liquid chromatography/ mass spectrometry
Biotransformation m/z Blood Hepatocytes Liver S9 Fraction
Mouse RatMonkeyHuman Mouse RatMonkeyHuman Mouse RatMonkeyHuman
O-Demethylation, O-dealkylation 107.0703bNDNDND ND ND ND ND ND NDND ND ND
O-Demethylation, O-dealkylation 151.0965bNDNDND ND ND ND ND ND NDND ND ND
O-Demethylation, O-dealkylation 195.1227bNDNDND ND ND ND ND ND NDND ND ND
Hydrolysis,N-Dealkylation 214.2529bNDNDND ND ND ND ND ND NDND ND ND
N-Dealkylation, oxidation 227.2017aNDNDND ND ND ND ND ND NDND ND ND
Hydrolysis (amine) 410.4720b++ND ND + + + + ++ + +
N,N-Didealkylation 531.5849bNDNDND ND ND ND ND ND NDND ND ND
N-Dealkylation 580.6396bNDNDND ND ND ND ND ND NDND ND ND
O-Demethylation, oxidation 629.6853bNDNDND ND ND ND ND ND NDND ND ND
Hydroxylation 633.6931bNDNDND ND ND ND ND ND NDND ND ND
ω-Hydroxylation, Oxidation 637.1880bNDNDND ND ND ND ND ND NDND ND ND
Hydrolysis (acid) 708.7721bNDNDND ND ND ND ND ND NDND ND ND
Note: Both theoretical and observed metabolites are included.
m/z = mass to charge ratio; ND = Not detected; + = metabolite present.
a. Negative ion mode .
b. Positive ion mode.
090177e196137069\Approved\Approved On: 21-Jan-2021 23:22 (GMT)
FDA-CBER-2021-5683-0013920