T-0016≪ TETRAHYDROISOQUINOLINE ALKALOIDAL COMPOUNDS WITH
MACROCYCLIC SCAFFOLD EXHIBITING ANTI-TUMOR ACTIVITIES≫＜Invention Overview＞■ Tetrahydroisoquinoline Alkaloids
Ecteinascidin743 (ET-743), a natural product isolated from a marine sponge, has been clinically applied as
an antitumor agent (Trabectedin, the trade name "Yondelis"). Structurally, ET-743 can be divided into 2 parts,
a core scaffold (unit A) consisting of two tetrahydroisoquinoline (THIQ) rings, and a 10-memberedmacrolactone ring with an additional THIQ moiety (unit B). Unit A recognizes DNA duplexes throughhydrogen bonds and alkylates at minor groove of DNA sequence-selectively. Unit B, meanwhile, is thought toinhibit the approach of nucleic proteins (e.g., transcription factor, DNA-repair proteins) to DNA, and thereby
inducing DNA double-strand break and cell death. A synthetic derivative, lurbinectedin (ZEPZELCA),
bearing a tetrahydro-β-carboline ring in place of the THIQ ring on unit B, has also been approved as achemotherapeutic agent for small cell lung cancer. Based on the structural and mechanistic insights into thepotent and unique anti-cancer agent ET-743, diversification of unit B is a promising approach to generate
novel antitumor agents targeting DNA–protein interactions. However, the commercialized semisynthetic
process of ET-743 requires more than 20-step chemical conversions starting from cyanosafracin B obtained
from microbial cultures, which makes it difficult to modify the structure of unit B.
Exploiting our experimental findings, we designed an unnatural, novel macrocyclic scaffold connecting C1and C5 hydroxyl group of unit A, instead of C4 position in ET-743. This design enabled rapid synthesis of aseries of drug lead candidates ranging from 14 to 17 membered macrocycles exhibiting comparable antitumor
activity with ET-743. This versatile synthetic platform with significantly fewer number of steps also allowed
flexible modification of unit B by just a single step appendage reaction (such as [4+2] cycloaddition).
Backgrounds

MeOHOHONH

MeO

MeNH
MeO MeO
Unit B
HO18SOAcO 5 4N MeMeN 21
Unit A18OOO 10 HOAcO O S HH4NMeNO1O21HOHHOMeMeHO 18
AcO O SHH4MeNMeN
8121OHO
H OH

nuclearprotein

DNAecteinascidin 743(Trabectedin, Yondelis®) : > 20 steps from cyanosafracin B

This invention
14MeN

Me15 NOMeMeHOO
MeO NH H H
54N

MeMeO

121HCNH

MeNO＜For inquiries, contact＞

MeNNOO

H

16 NH

CNH

HOMeO NH H H
5NNO

MeO

TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

OHCNMeH

HOMeO NH H H
5NNO

MeO

1OH

HP： https://todaitlo.com/R'4HNR

MeN
1OH

ET-743 type scaffold

Designed scaffold

17 NH

OMe
MeS

5OOOOOMeMeHOO
H
5 NH HO

10AcOlurbinectedin(ZEPZELCA®): < 10 steps from cyanosafracin B
R1NHNHOMeO

OMe
MeCNOMeMeO
5MeH

14 –17R’

4N

RN
1CNMe

T-0016■Evaluation of anti-cancer activity
These structurally simplified macrocyclic compounds can be synthesized within 10 steps in a time- andcost-efficient manner. Antitumor activities (GI50 values) of the synthetic compounds were evaluated against
39 cancer cell lines (JFCR39). Most of the macrocyclic compounds generated in this invention exhibit superior
antitumor activities compared to the natural product, cyanosafracin B and synthetic intermediates without
the macrocyclic scaffold. Notably, several synthetic compounds (e. g. TFR5) exhibited very potent antitumor
activity comparable to approved drugs, ET-743 and lurbinectedin.

Modification of macrocyclic structure (unit B) of ET-743 allowed development of the systematic collectionsof novel anti-tumor drug candidates. This synthetic platform rapidly generates more than 10 kinds ofskeletally diverse macrocyclic compounds capable of further structural modifications exploiting thepreinstalled functional groups (secondary amine, conjugated diene, and alkyne). The versatile and flexible
synthetic platform enables rational structural diversification of the complex macrocyclic structures, while
maintaining potent antitumor activities with inhibition of DNA–nuclear protein interactions.

＜Inventor＞
Professor, Hiroki Oguri, The University of Tokyo
Assistant Professor, Ryo Tanifuji, The University of Tokyo＜Public Information＞■Patent Applications： PCT/JP2022/008664＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Supplemental Data
Table1 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table2 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table3 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table4 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table5 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table6 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)

TFR5＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table7 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table8 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table9 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table10 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table11 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table12 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/

T-0016
Table13 :antitumor activity [nM] of Compound 29 (GI50 value: 50% growth inhibitory concentration)＜For inquiries, contact＞
TODAI TLO, Ltd. Haruno Wakayama
TEL：03-5805-7672 Email：wakayama@todaitlo.jp

HP： https://todaitlo.com/