Design and synthesis of novel

ᓫᇛ኱Ꮫ ༤ኈㄽᩥ

ᢠ

HBV

᪂つ

4 ′

Design and synthesis of novel

4′-modified nucleoside analogues as anti-HBV drugs

௧࿴ඖᖺᗘ

㨣ሯ ೺࿃

┠

┠ ḟ

➨㸯❶ ⥴ ㄽ

➨㸯⠇

B

➨㸯㡯

HBV

... ... 1

➨㸰㡯

HBV

... ... 2

➨㸱㡯

HBV

... ... 4

➨㸰⠇

HBV

HBV

... 6

➨㸰㡯 ⸆ ๣ ⪏ ᛶ

HBV... 8

➨㸱⠇

NAs

NAs

... 9

➨㸰㡯

4′-

NAs... 11

➨㸱㡯

4′-

NAs

... ... 14

➨㸲㡯 ᢠ

HBV

4′-

NAs... 15

➨㸰❶

4′ -

-5-

-4-

... ... 17

➨㸰⠇

4′ -࢔ ࢪ ࢻ -5-࣓ ࢳ ࣝ -4-⨨ ᥮ ࣆ ࣜ ࣑ ࢪ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ

... 19

➨㸱⠇

4′ -࢔ ࢪ ࢻ -5-࣓ ࢳ ࣝ -4-⨨ ᥮ ࣆ ࣜ ࣑ ࢪ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ

HBV

... 22

➨㸲⠇ ⪃ ᐹ

... .... 23

➨㸱❶

4′ -

... ... 24

➨㸰⠇

4′ -⨨ ᥮ ࢖ ࢯ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ ྜ ᡂ ࡟ 㛵 ࡍ ࡿ ᳨ ウ

4′-

... 27

➨㸰㡯

4′-

- 3′-

... ... 27

➨㸱㡯

4′-

... 30

➨㸲㡯

2-

- 4′-

... 33

➨㸳㡯

4′-

... ... 34

➨㸱⠇

4′ -⨨ ᥮ ࢖ ࢯ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ ᢠ ࢘ ࢖ ࣝ ࢫ ά ᛶ ཬ ࡧ

⣽ ⬊ ẘ ᛶ ホ ౯

... ... 35

➨㸲⠇ ⪃ ᐹ

... .. 36

➨㸲❶ ⥲ ᣓ

... .. 39

ㅰ ㎡

... ... 41

ᐇ 㦂 ࡢ 㒊

... . 43

ཧ ⪃ ᩥ ⊩

... 71

␎

␎ ㄒ ⾲

[A]

Ac : acetyl ADV : Adefovir

AIBN : 2,2 ′-azobis (isobutyronitrile)

APCI : atmospheric aressure chemical ionization

aq : aqua

AZT : 3′ -azido-3′ -dehydroxythymidine [B]

Bz : benzoyl

[C]

calcd. : calculated

cccDNA : covalently closed circular DNA CC

: 50% cytotoxic concentration CDI : 1,1 ′-carbonyldiimidazole [D]

dATP : 2′ -deoxy adenosine triphosphate DABCO : 1,4-diazabicyclo [2.2.2] octane DBU : 1,8-diazabicyclo [5.4.0] undec-7-ene DCM : dichloromethane

dCTP : 2′ -deoxy cytidine triphosphate ddC : 2′,3′-dideoxycytidine

ddI : 2′,3′-dideoxyinosine

DDQ : 2,3-dichloro-5,6-dicyano-p -benzoquinone dGTP : 2′ -deoxy guanosine triphosphate

DIAD : diisopropyl azodicarboxylate DMAP : N ,N-dimethyl-4-aminopyridine DMF : N ,N-dimethylformamide

DMP : Dess-Martin periodinane DMSO : dimethyl sulfoxide DMTr : 4,4 ′-dimethoxytrityl

d4T : 2′,3′-didehydro- 2′-deoxythymidine

[E]

EC

: half maximal (50%) effective concentration

EDC : 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide ESI : electrospray ionization

Et : ethyl

ETV : Entecavir [H]

HBe : hepatitis B virus envelope HBs : hepatitis B virus surface HBV : hepatitis B virus

HCV : hepatitis C virus

HIV : human immunodeficiency virus [I]

IFN : interferon

isoddA : 2′,3′-dideoxy isoadenosine [L]

LAM : Lamivudine LP : lone pair [M]

m CBA : m -chlorobenzoic acid

m CPBA : m -chloroperoxybenzoic acid Me : methyl

mRNA : messenger RNA MS 3A : molecular sieve 3

MS : mass spectrometry

MTT : 3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide [N]

Nap : (2-naphthalenyl) methyl NAs : nucleoside analogues n -BuLi : n -butyllithium

NMR : nuclear magnetic resonance nor-AZADO : 9-azanoradamantane-N -oxyl

NTCP : Na

-taurocholate cotransporting polypeptide

[P]

PCR : polymerase chain reaction pgRNA : pregenomicRNA

PMB : p -methoxybenzyl PPh

: triphenylphosphine PTSA : p -toluenesulfonic acid [Q]

QOL : quality of life [R]

rcDNA : relaxed circular DNA [T]

TAF : Tenofovir alafenamide

TBAF : tetrabutylammonium fluoride

TBA-HS : tetrabutylammonium hydrogen sulfate TBDPS : tert -butyldiphenylsilyl

TBS : tert -butyldimethylsilyl

TCDI : 1,1 ′-thiocarbonyldiimidazole TCID

: tissue culture infectious dose 50%

TDF : Tenofovir disoproxyl TEA : triethylamine

TFA : trifluoroacetic acid THF : tetrahydrofuran TIPS : triisopropylsilyl

TTP : thymidine triphosphate

1

➨

1❶ ⥴

➨

1⠇ B

➨

1

HBV

HBV (B

)

2

5,700

,

1%

ࡿ

.

HBV

,

,

㐍 ᒎ ࡍ ࡿ ࡇ ࡜ ࡀ ▱ ࡽ ࢀ

, 2010

HBV

.

, 2015

887,000

ே ࡀ

B

.

,

࢔ ࣇ ࣜ ࢝ ࡸ ༡ ⡿

,

, HBV

.

HBV

,

,

࡟ ࡼ ࡿ Ỉ ᖹ ឤ ᰁ ࡀ ୺ ࡞ ཎ ᅉ ࡜ ࡞ ࡿ

.

ᮏ 㑥 ࡛ ࡣ

,

1985

B

,

.

HBV

, HBV

.

,

,

A

HBV

.

ࡼ ࠺ ࡞ ஦ ែ ࢆ ៧ ៖ ࡋ

, WHO

HBV

࣮ ࢧ ࣝ ࣡ ࢡ ࢳ ࢿ ࣮ ࢩ ࣙ ࣥ ࠖ ࢆ ⾜ ࠺ ࡼ ࠺ ࡟ ່ ࿌ ࡋ ࡚ ࠾ ࡾ

,

ࡿ

.

,

.

ࡑ ࡢ ࡓ ࡵ

, HBV

,

☜ ❧ 㺃ᨵ Ⰻ ࢆ ⾜ ࠺ ࡇ ࡜ ࡣ ឤ ᰁ ⪅ ࡢ

QOL

.

2

➨

2

HBV

HBV

,

,

.

,

㉳ ࡇ ࡍ ࡇ ࡜ ࡀ ࠶ ࡾ

,

,

70-80%

.

,

࡜ ࡞ ࡗ ࡓ ሙ ྜ

,

,

.

,

95%, 1-5

20-30%,

5%

.

ࣜ ࢔ ࡜ ࡞ ࡿ

.

.

,

.

ࢃ ࡿ ࡜ ⫢ ⅖ ࡢ ⑕ ≧ ࢆ ࿊ ࡋ ጞ ࡵ ࡿ

.

⟅ ᮇ

,

.

HBV

࣮ ࢝ ࣮ ࡢ ≧ ែ ࡛ ༊ ู ࡉ ࢀ ࡿ

.

,

,

ࢆ ࿊ ࡉ ࡞ ࡃ ࡞ ࡗ ࡓ ᮇ 㛫 ࢆ մ ᐶ ゎ ᮇ ࡜ ࿧ ࡪ

. (Fig.1 )

ᣢ ⥆ ឤ ᰁ ࡢ ≧ ែ ࡣ ኱ ࡁ ࡃ ศ ࡅ ࡚ ࡇ ࡢ

4

,

HBe

, HBe

,

3

HBs

, HBs

.

Table 1

࡜ ࡵ ࡓ

.

HBe

HBV

. HBe

ྜ

, HBV

,

,

.

, HBe

, HBV

.

ࡲ ࡓ

, HBs

HBV

.

, HBs

,

HBV

, HBs

,

HBV

.

, HBe

, HBs

. HBe

HBV

,

, HBs

HBV

⋓ ᚓ ࡋ ࡚ ࠸ ࡿ ࡇ ࡜ ࢆ ⾲ ࡍ

.

ࡑ ࡋ ࡚

, HBe

HBe

. HBe

HBe

, HBe

HBe

.

.

, HBe

HBV

,

⫢ ⑓ ኚ ࡀ 㐍 ⾜ ࡋ ࡚ ࠸ ࡿ ࡇ ࡜ ࢆ ♧ ၀ ࡍ ࡿ

.

,

ࣟ ࢥ ࣥ ࣂ ࣮ ࢪ ࣙ ࣥ ࡀ ࠸ ࠿ ࡟ ᪩ ࡃ ⏕ ࡌ ࡿ ࠿ ࡀ ண ᚋ ࢆ ᨵ ၿ ࡍ ࡿ ୖ ࡛

,

.

ࡍ ࡞ ࢃ ࡕ

, HBV

,

,

QOL

.

4

➨

3

HBV

⌧ ᅾ

, HBV

NAs (

)

IFN (

ࣟ ࣥ

)

.

IFN

.

, IFN

,

⏝ ࡀ ከ ᒱ ࡟ ࢃ ࡓ ࡾ

,

.

,

,

࠸ ࡓ

.

, HIV (ࣄ ࢺ ච ␿ ୙ ඲ ࢘ ࢖ ࣝ ࢫ )

NAs

LAM (1)

HBV

,

2000

LAM

.

, LAM

࡛ ࠶ ࡾ

,

,

.

ࡑ ࡢ ᚋ

, ADV ( 2)

2004

, ADV

LAM

࡚ ࡶ ᭷ ຠ ࡛ ࠶ ࡗ ࡓ

.

LAM

, LAM

ADV

,

HBV

,

.

, ADV

⭈ 㞀 ᐖ ࡀ ☜ ㄆ ࡉ ࢀ ࡚ ࠾ ࡾ

,

,

.

, 2006

ETV (3 )

⏝ ࡀ 㛤 ጞ ࡉ ࢀ ࡓ

. ETV

HBV

, 0.5 mg/day

࠺ ᴟ ᚤ 㔞 ࡛ ᙉ ຊ ࡟

HBV

(LAM:100 mg/day, ADV:10 mg/day).

, ETV

,

࠸ ࠺ ඃ ࢀ ࡓ ≉ 㛗 ࡀ ࠶ ࡿ

.

,

ETV

HBV

,

LAM

, LAM

, ETV

.

, TDF (4 )

2014

, TDF

, 300 mg/day

.

, Tenofovir

TAF (5)

, 2017

. TAF

TDF

࡟ ྲྀ ࡾ ㎸ ࡲ ࢀ ࡿ ࡓ ࡵ

, 25 mg/day

TDF

.

,

࡚ ࠸ ࡿ

. (Fig.2)

5

᭱ ㏆ ሗ ࿌ ࡉ ࢀ ࡓ ᪂ つ

NAs

,

,

Besifovir dipivoxil ( 6 )

2019

.

ࡽ ࡟

, NAs

IFN

HBV

ࡿ

.

NTCP

Dane

Myrcludex-

B

2019

Phase2

.

, HBV

ࣉ ࢩ ࢻ 㔜 ྜ ࢆ 㜼 ᐖ ࡍ ࡿ

Morphothiadine (7 )

HBV DNA

Nitazoxanide ( 8)

2019

Phase2

ࡿ

. (Fig.3)

6

➨

2⠇ HBV

➨

1㡯 HBV

HBV

, HBV

Dane

࿧ ࡤ ࢀ ࡚ ࠸ ࡿ

. Dane

3.2kbp

DNA (relaxed circular DNA, rcDNA)

,

ࣟ ࣮ ࣉ ࡀ そ ࡗ ࡓ ᵓ 㐀 ࢆ ࡋ ࡚ ࠸ ࡿ

.

HBV

1964

, HBV

࡞ ࠿ ࡗ ࡓ ࡓ ࡵ

,

.

, 2012

HBV

⬊ ࡟ ౵ ධ ࡋ

,

NTCP (Na

ࣥ ࢫ ࣏ ࣮ ࢱ ࣮

)ࠖ ࡀ Ⓨ ぢ ࡉ ࢀ ࡓ .

, NTCP

ᰴ ࡀ ᶞ ❧ ࡉ ࢀ

,

HBV

HBV

, HBV

.

HBV

, Dane

⢏ Ꮚ ࡣ

NTCP

( Fig.4,

),

( Fig.4,

).

,

⤒ ࡚

rcDNA

(Fig.4,

).

rcDNA

୺ ࡢ ᣢ ࡘ 㓝 ⣲ ࡟ ࡼ ࡗ ࡚

covalently closed circular DNA (cccDNA)

7

㠀 ᖖ ࡟ Ᏻ ᐃ ࡞

DNA

( Fig.4,

).

, cccDNA

DNA

RNA

3.5 kb

pgRNA

mRNA

( Fig.4,

).

cccDNA

, pgRNA

࡟ ຍ ࠼

, 3

mRNA

, mRNA

࢘ ࢖ ࣝ ࢫ ⏤ ᮶ ࡢ ࢱ ࣥ ࣃ ࢡ ㉁ ࡀ ⩻ ヂ ࡉ ࢀ ࡿ

(Fig.4,

).

, pgRNA

( Fig.4,

),

rcDNA

(Fig.4,

). rcDNA

ࡴ ࢾ ࢡ ࣞ ࢜ ࢝ ࣉ ࢩ ࢻ ࡣ

,

Dane

( Fig.4 ,

).

cccDNA

( Fig.4 ,

-չ )

HBV

.

, B

cccDNA

, cccDNA

,

⒪ ἲ ࡛ ࡣ

HBV

.

8

➨

2

HBV

๓ 㡯 ࡛ ♧ ࡋ ࡓ ࡼ ࠺ ࡟

HBV

,

࠸

.

,

,

, NAs

. NAs

HBV

෗ ࡢ 㐣 ⛬ ࢆ 㜼 ᐖ ࡍ ࡿ ࡇ ࡜ ࡛ ᢠ

HBV

,

HBV

ࢆ ᢚ ไ ࡍ ࡿ ࡶ ࡢ ࡛ ࠶ ࡾ

,

.

, NAs

,

HBV

,

.

NAs

,

HBV

.

࡞ ࡀ ࡽ

,

,

,

.

⸆ ๣ ⪏ ᛶ ⋓ ᚓ ࡢ ᶵ ᗎ ࡟ 㛵 ࡍ ࡿ ◊ ✲ ࡣ ┒ ࢇ ࡟ ࠾ ࡇ ࡞ ࢃ ࢀ ࡚ ࠾ ࡾ

, NAs

.

[ 4 6] ᭱ ࡶ 㧗 ࠸ 㢖 ᗘ ࡛ ⏕ ࡌ ࡿ

LAM

204

ࢽ ࣥ ࡀ ࣂ ࣜ ࣥ ࡶ ࡋ ࡃ ࡣ ࢖ ࢯ ࣟ ࢖ ࢩ ࣥ ࡟ ኚ ␗ ࡋ

,

180

.

, LAM

ETV

࡟ ᑐ ࡍ ࡿ ឤ ཷ ᛶ ࡀ ప ୗ ࡋ

,

184

, 202

ࣜ ࣥ ࡶ ࡋ ࡃ ࡣ

250

ETV

⪏ ᛶ ࢆ ⋓ ᚓ ࡍ ࡿ ࡇ ࡜ ࡀ ሗ ࿌ ࡉ ࢀ ࡚ ࠸ ࡿ

.

, TDF

TAF

, LAM

ETV

TDF

.

ࡇ ࡢ ࡼ ࠺ ࡟

,

, HBV

.

,

,

,

HBV

࡚ ⸆ ๣ ⪏ ᛶ ࡢ ⋓ ᚓ ࢆ チ ࡉ ࡎ

,

NAs

ࡿ

.

9

➨

3⠇ NAs

➨

1㡯 ⌧

NAs

NAs

HBV

pgRNA

rcDNA

㏫ ㌿ ෗ ࢆ 㜼 ᐖ ࡍ ࡿ

.

,

㏫ ㌿ ෗ ࢆ 㜼 ᐖ ࡍ ࡿ ࡇ ࡜ ࡛ ṇ ᖖ ࡞ ⣽ ⬊ ࡢ ά ື ࢆ ᐖ ࡍ ࡿ ࡇ ࡜ ࡞ ࡃ

, HBV

ࣇ ࢧ ࢖ ࢡ ࣝ ࡢ ࡳ ࢆ 㜼 ᐖ ࡛ ࡁ ࡿ

.

,

DNA

࣮ ࢮ ࡜ ẚ ㍑ ࡋ ࡚ ᇶ ㉁ ≉ ␗ ᛶ ࡀ ప ࠸ ࡇ ࡜ ࡀ ▱ ࡽ ࢀ ࡚ ࠸ ࡿ

.

,

㏫ ㌿ ෗ 㓝 ⣲ ࡀ ࢾ ࢡ ࣞ ࢜ ࢳ ࢻ ࡟ 㢮 ఝ ࡋ ࡚ ࠸ ࡿ ໬ ྜ ≀ ࢆ ㄗ ࡗ ࡚ ྲྀ ࡾ ㎸ ࡳ

, DNA

.

NAs

(

)

.

NAs

dATP, dGTP, dCTP, TTP

࠿ ࡜ ➇ ྜ ࡋ ࡚ ㏫ ㌿ ෗ 㓝 ⣲ ࡟ ྲྀ ࡾ ㎸ ࡲ ࢀ

, HBV DNA

. DNA

⤌ ࡳ ㎸ ࡲ ࢀ ࡓ

NAs

5 ′

3 ′

DNA

. (Fig.5 )

ࡇ ࡢ ࡼ ࠺ ࡟ ㏫ ㌿ ෗ 㓝 ⣲ ࢆ 㜼 ᐖ ࡋ ࡚ ᢠ ࢘ ࢖ ࣝ ࢫ స ⏝ ࢆ ♧ ࡍ

NAs

, HIV

10

⒪ ࡢ ศ 㔝 ࡟ ࠾ ࠸ ࡚ ኱ ࡁ ࡃ ◊ ✲ 㺃㛤 Ⓨ ࡀ 㐍 ᒎ ࡋ ࡚ ࡁ ࡓ

. 1985

,

⑓ ࡛ ࠶ ࡗ ࡓ

HIV

NAs

AZT ( 9)

, HIV

.

, AZT

NAs

, HIV

. HIV

NAs

Fig.6

.

᭱ ึ ࡟ 㛤 Ⓨ ࡉ ࢀ ࡓ

AZT

3 ′఩ ࡢ Ỉ 㓟 ᇶ ࢆ ࢔ ࢪ ࢻ ᇶ ࡟ ⨨ ᥮ ࡋ

NAs

. AZT

HIV

DNA

, TTP

➇ ྜ ࡋ ࡚ ㏫ ㌿ ෗ 㓝 ⣲ ࡟ ྲྀ ࡾ ㎸ ࡲ ࢀ

, HIV

DNA

.

,

, 3′

DNA

,

㏫ ㌿ ෗ ࡀ 㜼 ᐖ ࡉ ࢀ ࡿ

.

AZT

NAs

ࡽ ࢀ ࡚ ࠾ ࡾ

, ddI (10 )

2′

3′఩ ࡢ Ỉ 㓟 ᇶ ࡀ Ỉ ⣲ ࡟ ⨨ ᥮ ࡉ ࢀ

NAs

. ddI

,

,

࡟ ⣽ ⬊ ෆ ࡢ 㓝 ⣲ ࡟ ࡼ ࡗ ࡚ ࣉ ࣜ ࣥ

6

ddATP

ࡿ

. ddATP

dATP

, AZT

DNA

.

, ddC (11 )

ࣥ ࡢ

3′఩ ࡢ Ỉ 㓟 ᇶ ࢆ Ỉ ⣲ ࡟ ⨨ ᥮ ࡋ ࡓ NAs

,

dCTP

, DNA

.

2′,3 ′-endo-olefin

᭷ ࡍ ࡿ

d4T (12 )

TTP

, DNA

.

ࡲ ࡓ

,

HBV

NAs

LAM

ADV, TDF, TAF

3′

, DNA

ࢆ ᢚ ไ ࡋ

,

.

, ETV

3 ′఩ ࡟ ┦ ᙜ ࡍ ࡿ ఩ ⨨ ࡢ

. ETV

㏫ ㌿ ෗ ࡟ ࡼ ࡗ ࡚

HBV

DNA

,

, DNA

.

ETV

DNA

,

㌿ ෗ 㓝 ⣲ ࡀ ㄆ ㆑ ࡛ ࡁ ࡞ ࡃ ࡞ ࡿ ࡇ ࡜ ࡟ ㉳ ᅉ ࡍ ࡿ ࡜ ⪃ ࠼ ࡽ ࢀ ࡿ

.

11

ࡇ ࡢ

ETV

3 ′

NAs

, 4 ′-

NAs

. 4′-

NAs

,

࢖ ࣝ ࢫ స ⏝ ࢆ ♧ ࡋ

,

.

➨

2

4 ′-

NAs

4 ′-⨨ ᥮ NAs

4 ′఩ ࡟ ࢚ ࢳ ࢽ ࣝ ᇶ ,

. ( Fig.7)

,

HIV

⏝ ࡉ ࢀ ࡚ ࠸ ࡿ

Censavudine ( 13)

4′

4′-

NAs

࠶ ࡿ

.

,

Phase2

Islatravir (14)

4′఩

࡟ ࢚ ࢳ ࢽ ࣝ ᇶ ࢆ ᭷ ࡍ ࡿ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡛ ࠶ ࡾ

, 3′఩ ࡟ Ỉ 㓟 ᇶ ࢆ ᣢ ࡘ ࡟ ࡶ ࠿ ࠿

ࢃ ࡽ ࡎ

,

. 4′-

NAs

1956

Streptomyces calvus

Nucleocidin ( 15 )

.

1971

Moffatt

16

4′,5′- exo-olefin

17

,

௜ ຍ ཯ ᛂ ࡟ ࡚

4 ′

18

,

15

. (Scheme 1)

12

ࡑ ࡢ ᚋ

, 1992

, Maag

4 ′఩ ࡟ ࢔ ࢪ ࢻ ᇶ ࢆ ᑟ ධ ࡋ ࡓ 4 ′-

NAs (19 -22 )

,

HIV

. (Table 2)

௨ 㝆

,

4′ -

NAs

,

, 4′

23

24 ,

25,

26 , 27

HIV

. (Fig.8)

4 ′-⨨ ᥮ NAs

3 ′఩ ࡟ Ỉ 㓟 ᇶ ࢆ

,

,

࡟ ㉳ ᅉ ࡍ ࡿ ࡜ ⪃ ࠼ ࡽ ࢀ ࡿ

.

13

14

➨

3

4 ′-

NAs

ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡟ ࠾ ࡅ ࡿ ⢾ ࡢ ❧ య 㓄 ᗙ ࡣ

C2 ′-endo/C3′- exo

(South

)

C2 ′- exo/C3 ′-endo

(North

)

,

࢔ ࢺ ࣜ ࢔ ࣝ ఩ ࡜ ࡞ ࡿ

South

.

, 4 ′

,

, North

. (Fig.9 )

4 ′-

NAs

North

,

4 ′

࢔ ࢟ ࢩ ࢔ ࣝ ఩ ࢆ ༨ ࡵ ࡿ ࡓ ࡵ

,

,

.

North

.

࣐ ࣮ ຠ ᯝ ࡣ

,

4′

᥮ ᇶ ࡢ

σ*

,

ࡿ ࡇ ࡜ ࡛ ⏕ ࡌ ࡿ

. (Fig.10 )

, 4′఩ ࡟ ❅ ⣲ ཎ Ꮚ ࡸ 㓟 ⣲ ཎ Ꮚ ,

ࣥ ࡞ ࡝ ࡀ ⤖ ྜ ࡋ ࡚ ࠸ ࡿ ሙ ྜ

,

North

࡞ ࡿ

.

15

ࡋ ࠿ ࡋ

,

.

, 4′

Islatravir

North

Kirby

.

⏤ ࡜ ࡋ ࡚ ࡣ ஬ ဨ ⎔ ࢆ ᙧ ᡂ ࡍ ࡿ 㓟 ⣲ ཬ ࡧ

, 3′఩ ࡢ 㓟 ⣲ ࡜ ࢚ ࢳ ࢽ ࣝ ᇶ ࡢ π*㌶ 㐨

. (Fig.11 )

ࡇ ࡢ ࡼ ࠺ ࡟

4′-

NAs

DNA

,

.

➨

4㡯 ᢠ

HBV

4′-

NAs

๓ 㡯 ࡛ ㏙ ࡭ ࡓ ࡼ ࠺ ࡟

, 4′-

NAs

HIV

,

࡛ ࠸ ࡿ ࡶ ࡢ ࡀ ࠶ ࡿ

.

, HIV

HBV

4 ′-⨨ ᥮ NAs

.

,

࡛ ࠶ ࡿ ‶ ᒇ ᩍ ᤵ ࡢ ࢢ ࣝ ࣮ ࣉ ࡟ ࡼ ࡾ

,

4 ′-⨨ ᥮ NAs

HBV

.

HBV

4′-

NAs

Fig.12

.

16

ࣉ ࣜ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ

4 ′఩ ࡟ ࢩ ࢔ ࣀ ᇶ ࢆ ᑟ ධ ࡋ ࡓ CdG ( 28 )

CAdA (29 )

HBV

(EC

= 0.0004 μ M)

, ETV

࡟ ᑐ ࡋ ࡚ ࡶ ᙉ ࠸ ᢠ

HBV

(EC

= 0.04-0.07 μ M)

.

, ETV

4′఩ ࡟ ࢩ ࢔ ࣀ ᇶ ࢆ ᑟ ධ ࡋ ࡓ CMCdG ( 30 )

ETV

HBV

(ETV:EC

= 64.037 μ M, CMCdG:EC

= 2.657 μ M)

࠶ ࡿ

.

,

NAs

(CC

= 2.6-89 μ M)

.

ࡇ ࡢ ࡼ ࠺ ࡟

4 ′-⨨ ᥮ NAs

HBV

,

࡚ ࡶ ຠ ᯝ ࡀ ࠶ ࡿ ࡇ ࡜ ࡀ ♧ ࡉ ࢀ ࡓ

.

HBV

,

, HBV

CdG

, A87, F88, P177, L180, M204, D205

㎸ ࡳ

,

.

4 ′-

NAs

,

HBV

ࡽ ࡎ

,

.

,

HBV

,

NAs

,

4 ′-⨨ ᥮ NAs

,

.

17

➨

2

4 ′-

-5-

-4-

➨

1⠇ ᗎ

ඛ ㏙ ࡋ ࡓ ࡼ ࠺ ࡟

4 ′-

HBV

.

4 ′-

࢜ ࢩ ࢻ ࡛ ࠶ ࡿ

AdG (31)

AAdA (32 )

HBV

(EC

= 0.006-0.009 μ M)

.

, AdG

AAdA

(CC

= 4.6-4.9 μ M),

4 ′-

. (Fig.13 )

ࡑ ࡇ ࡛

,

.

HBV

἞ ⒪ ⸆ ࡜ ࡋ ࡚ ౑ ⏝ ࡉ ࢀ ࡚ ࠸ ࡿ

Telbivudine (33)

.

,

4′-

(34 )

࠸ ࡚ ᢠ

HBV

.

,

5-

FddMeC (35 )

HBV

(EC

= 0.54 μ M)

࡜ ࡀ ሗ ࿌ ࡉ ࢀ ࡚ ࠸ ࡓ

. (Fig.14)

ࡲ ࡓ

,

HBV

FNC (36)

, N

NAs ( 37- 38)

HBV

.

( Fig.15)

18

௨ ୖ ࡢ ▱ ぢ ࢆ ࡩ ࡲ ࠼

,

4′-

( 34 )

, 5-

4′-

-2 ′-

-5-

( 39)

HBV

,

HBV

࡜ ⪃ ࠼ ࡓ

.

, N

/ O

,

ࢆ ᮇ ᚅ ࡋ ࡚

, NAs 40 -44

.

, 4′-࢔ ࢪ ࢻ -5-࣓ ࢳ ࣝ -4-⨨ ᥮ ࣆ

ࣜ ࣑ ࢪ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ

(34 , 39 -44 )

HBV

. (Fig.16 )

19

➨

2

4 ′-

- 5-

-4-

34

Maag

. (Scheme 2)

ࢳ ࣑ ࢪ ࣥ ࢆ ฟ Ⓨ ཎ ᩱ ࡜ ࡋ ࡚

,

5′఩ ࡢ Ỉ 㓟 ᇶ ࢆ

ࣚ ࢘ ⣲ ࡟ ⨨ ᥮ ࡋ ࡓ

45

, E2

, 4 ′,5′-exo-olefin

46

. 46

47

, 3 ′

p-

48

. 48

ࢆ

mCPBA

, p-

,

3-ࢡ ࣟ

ࣟ Ᏻ ᜥ 㤶 㓟 ࢔ ࢽ ࢜ ࣥ ࡢ ồ ᰾ ⨨ ᥮ ཯ ᛂ ࡟ ࡼ ࡾ

49

.

, 49

7M

34

.

ࡉ ࡽ ࡟

,

4

, 49

4

໬ ࡋ ࡓ 㘽 ୰ 㛫 య

50

. (Scheme 3)

20

㘽 ୰ 㛫 య

50

4

DABCO

,

ࣝ ࡟ ࡼ ࡿ

S

Ar

,

40- 41

. (Scheme 4 )

ࡲ ࡓ

,

50

, S

Ar

39 , 42 -43

.

,

50

4

DABCO

,

S

Ar

44

ࢆ ᚓ ࡓ

. (Scheme 5)

21

ḟ ⠇ ࡟ ࡚ ᢠ

HBV

.

22

➨

3

4 ′-

- 5-

-4-

HBV

ᢠ

HBV

,

.

ྛ ໬ ྜ ≀ ࡢ ᢠ

HBV

real-time PCR

.

96 well plate

, HepG2 2.2.15

1 well

4

10

,

200 μ L

.

,

ࣟ ࣮ ࣝ ࡜ ࡋ ࡚ ໬ ྜ ≀ ⁐ ᾮ ࢆ ຍ ࠼ ࡞ ࠸

well

. HepG2 2.2.15

HBV

, in vitro

HBV

.

well plate

37

CO

14

.

, 3

7

,

ᩚ ࡋ ࡓ ྛ ໬ ྜ ≀ ࢆ ྵ ࡴ ᾮ య ᇵ ᆅ ࡜ ஺ ᥮ ࡋ ࡓ

.

,

, HBV DNA

.

,

PCR

well

HBV DNA

,

DNA

EC

.

୍ ᪉

,

96 well plate

MT-2

1 well

2

10

,ྛ well

200 μ L

.

, Cell Counting Kit-8

CC

ྜ ᡂ ࡋ ࡓ

34, 39 -44

ETV

HBV

Table 3

.

᭱ ࡶ 㧗 ࠸ ᢠ

HBV

34

, EC

= 0.63s0.29

23

μ M

.

, 39

HBV

, EC

= 5.99

μ M

.

34

39

(CC

> 100 μM).

୍ ᪉

, N

/ O

40 -44

,

HBV

.

ḟ ⠇ ࡟ ࡚ ⤖ ᯝ ࡟ ࡘ ࠸ ࡚ ⪃ ᐹ ࡍ ࡿ

.

➨

4⠇ ⪃

ᮏ ❶ ࡛ ࡣ

,

HBV

NAs

4′-

-5-

-4-

᥮ ࣆ ࣜ ࣑ ࢪ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࢆ タ ィ ࡋ

,

.

4 ′-࢔ ࢪ ࢻ ࢳ ࣑ ࢪ ࣥ ࡢ ྜ ᡂ ࢆ 6

,

22.0%࡛ 㐩 ᡂ ࡋ ࡓ .

, 4′-

࢔ ࢪ ࢻ

-5-

-4-⨨ ᥮ ࣆ ࣜ ࣑ ࢪ ࣥ ࢾ ࢡ ࣞ ࢜ ࢩ ࢻ ࡢ ྜ ᡂ ࡛ 㔜 せ ࡞ 㘽 ୰ 㛫 య 50

6

,

17.3%࡛ 㐩 ᡂ ࡋ ࡓ .

, S

Ar

4

,

21.0-82.1%

.

ᮏ ❶ ࡛ タ ィ ࡋ ࡓ ໬ ྜ ≀ ࡛ ᭱ ࡶ 㧗 ࠸ ᢠ

HBV

34

ࡾ

(EC

= 0.63

μ M), 39

HBV

(EC

= 5.99

5.54 μ M).

, 34

39

(CC

> 100 μM)

34

39

࠼ ࡽ ࢀ ࡿ

.

, 4′-

(CC

= 0.21 μ M)

,

5-

࠶ ࡿ

39

,

HBV

.

,

ࣆ ࣜ ࣑ ࢪ ࣥ

5

.

, FNC

N

,

HBV

, 40- 44

HBV

.

,

5

,

N

/ O

ࣝ ໬ ࡋ ࡓ ሙ ྜ

,

HBV

.

,

HBV

4′-

NAs

࠾ ࠸ ࡚

,

5-

ࡽ ࢀ ࡿ

.

ࡇ ࡢ ࡇ ࡜ ࡣ ௒ ᚋ

,

HBV

4′-

NAs

ୖ ࡛ 㠀 ᖖ ࡟ ᭷ ┈ ࡞ ᝟ ሗ ࡛ ࠶ ࡿ

.