marine biodiversity what is it good for ? david j. newman, d.phil. chief, natural products branch...
TRANSCRIPT
Marine BiodiversityWhat Is It Good For ?
David J. Newman, D.Phil.
Chief, Natural Products BranchDevelopmental Therapeutics Program
Division of Cancer Treatment & DiagnosisNCI-Frederick,
Frederick, MD, 21702, USA
+1.301.846.5397 Voice+1.301.846.6178 Facsimile
Email [email protected]
Why Look There ?
A question that is very frequently asked is the title of this short
talk, and what I intend to try to do in the next 15 or so minutes
is to show you what is going on from the perspective of drug
discovery in utilization of the marine resources, meaning coral reefs,
marine muds and sessile invertebrates, but the emphasis as will
become apparent is on the organisms we cannot see.
Because of my background, the focus is on cancer, but any of the
materials may (will ?) have utility in other diseases.
O O
HOOAc
H3COOC
OOH
O
O COOCH3
O
O
OH
HO
Bryostatin 1 -Complex polyketide natural product-Not very effective by itself in Phase II trials-Was in Phase I and Phase II trials with other cytotoxic drug therapy (e.g. Vincristine) to melanoma, kidney cancer and lymphoma
Biosynthetic Source?
Pettit,GR Fortschritte der Chemie organischer Naturstoffe. 1991, 57, 153-
195.
Bryostatin 1 Modulates Protein Kinase C
Bryozoan-Bacterial Symbiosis Produces the Anticancer Bryostatins
(Haygood/Sherman)
• Not yet cultivated• Found in almost all
tested populations, adults and larvae (by PCR)
• Symbiont is a novel gamma
proteobacterium (by 16S rRNA)
– named “Candidatus Endobugula sertula”
• Pathway cloned with Sherman Lab• Many novel biosynthetic aspects
In situ hybridization with symbiont specific probe in larva
Haygood et al, Chem Biol 2005, 12, 397Sherman et al, J. Nat Prod 2007, 70, 67
Microbes and Tubulin Interactive Agents
Tubulin as a target has a relatively long history, though not with agents
from microbes (though just wait until a little later).
With the discovery of the mechanism of action of Taxol® by Susan
Horwitz in 1979, a new target came into play for antitumor agents.
The marine environment in particular has yielded some extremely
interesting molecules that interact with tubulin in a variety of ways
and that probably also involve microbes in their biosynthesis.
Potentially Microbial
OO
OH
HO
OH
OH O O
NH2
Discodermolide
HO
OH
OH O O
NH2
OO
DD4
CH3
O
CO2Me
H
OH
Me Me
OH
H
N
N
O
Me
Me
CH3
O
H
OH
Me Me
OH
H
N
N
O
Me
Me
AcO
O
O
OH
OHCH3
O
COOCH3
H
OCH3
H3C CH3
OH
H
N
NO
CH3
Sarcodicytin [Sarcodictyon roseum] Eleutherobin [Eleutherobia sp.]
Sarcorobin or Eleutherdyctin
Some Microbial Involvement ?
O
O O
O
HH
HOH
OH
H
HO
O
O O
O
HH
HO
HO
OH
H
Laulimalide Isolaulimalide
OHO
OO
O
HOO
O
HO
OH
OHOH
HO
O O
Dictyostatin
Peloruside A
OH
May well be a new binding site
Other Marine-Derived AgentsTubulin, Proteasome, VDA and
VoATPase
OHO
NH
N
OCl
N
O
ClHN
OHN
O
HO
HO
Original Diazonamide Structure
O X
NH
N
O
Cl
N
O
ClHNHN
O
HO
"OXO"-analogue, X = O
Revised Diazonamide Structure, X = NH
ON
NH
N
O
O
OH
O
E-7974
NH
OO
OOH
HH
Cl
H
Salinosporamide A
HN
NH
O
O
NH
N
NPI-2358
NH
O
OOH
OOHH
Salicylihalimide A
Me
CH2OH
H OH
HO H
Kishi Synthesis of Halichondrin B, 1992
• “Acyclic Stereochemistry Control”– Synthetic access to highly
complex natural products• New Ni/Cr-mediated coupling
reaction to form C-C bonds– Nozaki-Hiyama-Kishi reaction
• Why halichondrin B?– Showcase Ni/Cr-mediated
coupling– Highly potent anticancer
activity (Hirata & Uemura, 1986) was “added value”
• Halichondrin B total synthesis:– Aicher TD, Buszek KR, Fang
FG, Forsyth CJ, Jung SH, Kishi Y, Matelich MC, Scola PM, Spero DM, Yoon SK (1992) JACS 114:3162-3164
Professor Yoshito KishiDepartment of Chemistry
Harvard University
1,2,3-butane triolAsymmetric carbons: 2Possible stereoisomers: 2n = 22 = 4
Halichondrin BAsymmetric carbons: 32Possible stereoisomers: 2n = 232 = 4.3 x 109
O
H
H
O
O
H
H
O
O
H
HMe
O
O
MeH
H
O
O
Me
O
OO
O
OO
H
H
O
Me
HO
HHO
HOH
Total synthesis created an opportunity to develop halichondrin B-based drugs
from renewable resources
E7389: Synthetic Macrocyclic Ketone Analog of Halichondrin B’s “Right
Half”
O
Me
O
OO
O
OO
H
H
O
HO
OMe
H2N
OH
E7389
O
H
H
O
O
H
H
O
O
H
HMe
O
O
MeH
H
O
O
Me
O
OO
O
OO
H
H
O
Me
HO
HHO
HOH
Halichondrin B
~200 analogues
MW = 1110
32 stereocenters
0.2 nM (MDA-MB-435)
MW = 730
19 stereocenters
0.1 nM (MDA-MB-435)
Paclitaxel
MW = 854
11 stereocenters
2.5 nM (MDA-MB-435)
Eribulin, E7389 = NSC-707389previously ER-086526, B1939
“Piece de Resistance”; “Seabed to Sickbed”
N
N
O
O
O
O
H
OH
HO
O
H
H
NH
O
O
O
HO
S
Et743; Trabectedin; Yondelis(R)
The first “Direct from the Sea” drug
to be approved for Cancer Treatment.
EMEA 20SEP07
Just about every technique used; large-scale
harvesting, aquaculture and semisynthesis from
Cyanosafracin B
“You are what you eat”
Dolabella auricularia Dolastatins come from a Symploca species that they
graze on
N
HN
N
O
O
CH3 OCH3 O
N
OCH3 O
NH
N S
N
HN
N
O
O
CH3 OH O
N
OCH3 O
HN
Dolastatin 10
Dolastatin 10 and a Synthetic Analogue
Auristatin PEPhase I (II)
Marine Sediments: Nereus Pharmaceuticals Marine Microbe Culture Collection
Over 15,000 Strains~50 % Actinomycetes; 10 New genera discovered
~50 % Fungi
HN
O
O
Cl
OHHH
Salinosporamide A Fenical et al., Angew. Chem. Int. Ed., 42, 355-357 (2003)
Sediment sampler
DISCOVERY AND DEVELOPMENT OF NPI-0052, A NOVEL
PROTEASOME INHIBITOR FOR THE TREATEMENT OF CANCER
Feb 2003: NPI-0052 (Salinosporamide A) structure, cytotoxicity and proteasome inhibitory activity established (Feling et al)
2Q 2007:Phase I Multiple Myeloma
2Q 2006:Phase I Solid Tumors and Lymphoma
Mar 2006:X-Ray Crystal Structure in complex with 20S proteasome
Dec 2005:NPI-0052 efficacy in mouse multiple myeloma xenograft models (Chauhan et al)
Nov 2006:NPI-0052 efficacy in mouse colon cancer xenograft models (Cusack et al)
Dec 2005:NPI-0052 IND Filed
Preclinical Development (30 months)
Preclinical models (in vitro/in vivo)API manufacturing (saline fermentation)Formulation developmentDrug product manufacturingToxicology
May 2003:FDA approves Velcade™ for treatment of multiple myeloma, validating the proteasome as a target for cancer treatment
May 2004: Total synthesis (EJ Corey)
June 2005: Total synthesis (Danishefsky)
June 2007: Total synthesis (Ling et al)
Oct 2002:Novel Marine actinomycete Salinisporadiscovered (Mincer et al)
Salinosporamide Development Time Line
Ray Lam, Nereus Pharma
Imperial Purple and The Cell Cycle
Hexaplex trunculus (A) was extracted for Tyrian purple (B), various brominated indirubins (1–4) (C), and indigos. Oxime derivatives (5–8) and N1-methylated analogs (9–11) of these indirubins were synthesized, as well as the methoxime and acetoxime of 6-bromoindirubin (12, 13). 6-bromoindirubin-3-oxime (BIO) (7) and its control analog 1-methyl-6-bromoindirubin-3-oxime (MeBIO) (11) were used in the biological models.
Isolation of Indirubin-Binding Proteins
Potential Anti-Alzheimer’s Treatment
G2
M
G1
S
G0
tyrosine kinases
DNA synthesis
topoisomerase I
CDK2
tubulin polymerisation
/depolymerisati
on
Vinca alkaloidstaxol/taxoterehalichondrinspongistatinrhizoxincryptophycinsarcodictyin eleutherobinepothilonesdiscodermolideindibulindolastatincombretastatineribulin
camptothecin
CDK4
flavopiridol
(R)-roscovitine (CYC202)paullones, indirubins
gleeveciressaerlotinib
hydroxyureacytarabineantifolates
5-fluorouracil6-mercaptopurine
nitrogen mustardsnitrosoureasmitomycin C
CDK1
Chk1Chk2
UCN-01, SB-218078debromohymenialdisineisogranulatimide
AhR
actin
kinesin Eg5
monastrol
trabectedin
podophyllotoxin,doxorubicinetoposide, mitoxantrone
topoisomerase II
ATM/ATR
tipifarniblonafarnib
ROCK
Y27632
CDC25
DF203
FK317 HMGA
Plk1
Aurora
wortmannincaffeine
ODC/SAMDC
Pin1
GSK-3
Cdc7
nucleotide excision repair
Raf
cytochalasinslatrunculin Ascytophycinsdolastatin 11jaspamide
paullones, indirubins
(R)-roscovitine (CYC202)paullones, indirubins
sorafenib*
fumagillin,TNP-470PRIMA-1, pifithrin
rapamycin mTOR/FRAP
PS-341 proteasome
bryostatin, PKC412
PKC
histone deacetylase
trichostatin, FK228
HSP90
geldanamycin, 17-AAGATK, MAFP cytosolic phospholipase A2
hexadecylphosphocholine
phospholipase D
CT-2584 choline kinase
MEK1/Erk-1/2
PD98059, U0126
menadione (K3)
farnesyl transferase
phosphatases
okadaic acid, fostreicin, calyculin A
Wee1
PD0166285
polyamine analoguesPin1
p53/MDM2
Cell Cycle and Natural Products
Modified from Meijer, 2003