<110>    Azcuris Co., Ltd.

<120>    Screening Method of compound for IL-33 inhibiting and the
         Pharmaceutical composition comprising the same

<130>    OP20220028WO

<150>    KR 10-2021-0071014
<151>    2021-06-01

<160>    26

<170>    KoPatentIn 3.0

<210>    1
<211>    154
<212>    PRT
<213>    Homo sapiens

<400>    1
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Cys Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Asn Cys Val
            100                 105                 110 

Ser Phe Glu Cys Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Cys Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Leu Ser Glu Thr
145                 150                


<210>    2
<211>    462
<212>    DNA
<213>    Homo sapiens

<400>    2
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtgtgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactccaac tgtgtttcat ttgaatgcaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtgta ctgaaaatat cttgtttaag ctctctgaaa ct                           462


<210>    3
<211>    154
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) protein sequence


<400>    3
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Ser Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Asn Ala Val
            100                 105                 110 

Ser Phe Glu Ala Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Ser Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Leu Ser Glu Thr
145                 150                


<210>    4
<211>    462
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) DNA sequence


<400>    4
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtctgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactccaac gcggtttcat ttgaagcgaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtcta ctgaaaatat cttgtttaag ctctctgaaa ct                           462


<210>    5
<211>    154
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) N226C protein sequence


<400>    5
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Ser Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Cys Ala Val
            100                 105                 110 

Ser Phe Glu Ala Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Ser Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Leu Ser Glu Thr
145                 150                


<210>    6
<211>    462
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) N226C DNA sequence


<400>    6
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtctgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactcctgc gcggtttcat ttgaagcgaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtcta ctgaaaatat cttgtttaag ctctctgaaa ct                           462


<210>    7
<211>    154
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) V228C protein sequence


<400>    7
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Ser Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Asn Ala Cys
            100                 105                 110 

Ser Phe Glu Ala Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Ser Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Leu Ser Glu Thr
145                 150                


<210>    8
<211>    462
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) V228C DNA sequence


<400>    8
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtctgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactccaac gcgtgctcat ttgaagcgaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtcta ctgaaaatat cttgtttaag ctctctgaaa ct                           462


<210>    9
<211>    154
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) L267C protein sequence


<400>    9
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Ser Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Asn Ala Val
            100                 105                 110 

Ser Phe Glu Ala Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Ser Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Cys Ser Glu Thr
145                 150                


<210>    10
<211>    462
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) L267C DNA sequence


<400>    10
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtctgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactccaac gcggtttcat ttgaagcgaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtcta ctgaaaatat cttgtttaag tgctctgaaa ct                           462


<210>    11
<211>    154
<212>    PRT
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) E269C protein sequence


<400>    11
Ser Pro Ile Thr Glu Tyr Leu Ala Ser Leu Ser Thr Tyr Asn Asp Gln
  1               5                  10                  15 

Ser Ile Thr Phe Ala Leu Glu Asp Glu Ser Tyr Glu Ile Tyr Val Glu
             20                  25                  30 

Asp Leu Lys Lys Asp Glu Lys Lys Asp Lys Val Leu Leu Ser Tyr Tyr
         35                  40                  45 

Glu Ser Gln His Pro Ser Asn Glu Ser Gly Asp Gly Val Asp Gly Lys
     50                  55                  60 

Met Leu Met Val Thr Leu Ser Pro Thr Lys Asp Phe Trp Leu His Ala
 65                  70                  75                  80 

Asn Asn Lys Glu His Ser Val Glu Leu His Lys Ser Glu Lys Pro Leu
                 85                  90                  95 

Pro Asp Gln Ala Phe Phe Val Leu His Asn Met His Ser Asn Ala Val
            100                 105                 110 

Ser Phe Glu Ala Lys Thr Asp Pro Gly Val Phe Ile Gly Val Lys Asp
        115                 120                 125 

Asn His Leu Ala Leu Ile Lys Val Asp Ser Ser Glu Asn Leu Ser Thr
    130                 135                 140 

Glu Asn Ile Leu Phe Lys Leu Ser Cys Thr
145                 150                


<210>    12
<211>    462
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    IL-33 4Cysteine knock-out(K/O) E269C DNA sequence


<400>    12
ccacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt         60

gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa        120

gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt        180

gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc        240

aacaacaagg aacactctgt ggagctccat aagtctgaaa aaccactgcc agaccaggcc        300

ttctttgtcc ttcataatat gcactccaac gcggtttcat ttgaagcgaa gactgatcct        360

ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag        420

aatttgtcta ctgaaaatat cttgtttaag ctctcttgca ct                           462


<210>    13
<211>    27
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C208S_for primer


<400>    13
gagctccata agtctgaaaa accactg                                             27


<210>    14
<211>    27
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C208S_rev primer


<400>    14
cagtggtttt tcagacttat ggagctc                                             27


<210>    15
<211>    41
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C227A_C232A_for primer


<400>    15
atgcactcca acgcggtttc atttgaagcg aagactgatc c                             41


<210>    16
<211>    41
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C227A_C232A_rev primer


<400>    16
ggatcagtct tcgcttcaaa tgaaaccgcg ttggagtgca t                             41


<210>    17
<211>    35
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C259S_for primer


<400>    17
ctcttctgag aatttgtcta ctgaaaatat cttgt                                    35


<210>    18
<211>    35
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    C259S_rev primer


<400>    18
acaagatatt ttcagtagac aaattctcag aagag                                    35


<210>    19
<211>    37
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    L267C_for primer


<400>    19
gaaaatatct tgtttaagtg ctctgaaact tgagtcg                                  37


<210>    20
<211>    34
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    L267C_rev primer


<400>    20
gtcgactcaa gtttcagagc acttaaacaa gata                                     34


<210>    21
<211>    33
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    N226C_for primer


<400>    21
cataatatgc actcctgcgc ggtttcattt gaa                                      33


<210>    22
<211>    33
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    N226C_rev primer


<400>    22
ttcaaatgaa accgcgcagg agtgcatatt atg                                      33


<210>    23
<211>    28
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    V228C_for primer


<400>    23
atgcactcca acgcgtgctc atttgaat                                            28


<210>    24
<211>    29
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    V228C_rev primer


<400>    24
cttgcattca aatgagcacg cgttggagt                                           29


<210>    25
<211>    30
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    E269C_for primer


<400>    25
ttgtttaagc tctcttgcac ttgagtcgac                                          30


<210>    26
<211>    27
<212>    DNA
<213>    Artificial Sequence

<220>
<223>    E269C_rev primer


<400>    26
ctcgtcgact caagtgcaag agagctt                                             27
