英文文献汇报PhleghenrinesA

椭圆马尾杉Phleghenrines

A−D

and

Neophleghenrine

A,

Bioactive

and

StruRigid

Lycopodium

Alkaloids

from

Phlegmariurus

henryiReporter:

Lai

YunfeiDate:

2017/3/31半月刊、主要以有机化学为主（物理与理论化学、生物化学）organic

lettersPublication

periods

:SemimonthlyIF

:6.732Fields:Organic

Chemistry、Physical

anTheoretical

Chemistry、BiochemistryCONTENTS1Introduction2

Experimental3

Results

and

discussion4

HighlightIntroduction01阿尔兹海默症(AD)是一种常见的与年龄有关的慢性退化性脑疾病，公认为痴呆最常见的形式，在所有病例中占了60−80%。乙酰胆碱酯酶(AChE)能增加脑内神经递质乙酰胆碱的水平，仍然是AD的症状改善最有价值的治疗靶点。在由美国食品和药物管理局（FDA）批准的（截至2015十二月）的六种药物中，四种是乙酰胆碱酯酶抑制剂：加兰他敏，多奈哌齐，卡巴拉汀，和他克林。但是，这些药物只是暂时改善，而不是治愈，AD的症状，并有常见的胃肠道副作用，如恶心和呕吐。因此，当务之急是开发新的药物治疗，以减缓或停止AD的进展。Alzheimer’s

disease

(AD)

is

a

common

age-related,

chronic

degenerative

brain

disease

and

recognized

as

the

most

common

form

of

dementia,

accounting

for

an

estimated

60−80%

of

all

cases.Among

the

six

drugs

approved

(as

of

December

2015)

by

the

U.S.

Food

and

Drug

Administration

(FDA),

four

are

AChE

inhibitors:

galantamine,

rivastigmine,

donepezil,

and

tacrine.However,

each

of

these

drugs

only

temporarily

improves,

rather

than

cures,

symptoms

of

AD

and

has

common

gastrointestinal

side

effects

such

as

nausea

and

vomiting.galantaminerivastigminetacrinedonepezil石松生物碱是从石松科植物和其近缘植物分离得到天然化合物，具有广泛的生物活性，包括乙酰胆碱酯酶抑制活性。这类生物碱生物碱有被分成四类结构：lycopodine（石松碱）型，lycodines（石松定）型，fawcettimine（法氏石松碱）型和其他。石杉碱甲和石杉碱乙，在1986从蛇足石杉中首次分离，是lycodine型石松生物碱代表物，表现出对乙酰胆碱酯酶有效的抑制活性。石杉碱甲已在中国作为一种治疗AD的新药上市,石杉碱甲（HupA）在治疗创伤性脑损伤（TBI）的影响的第二阶段临床试验也在进行中。fawcettimines,

and

the

miscellaneous.Lycopodium

alkaloids

are

from

the

genus

Lycopodiu➢

Huperzines

A

and

B

,

first

isolated

m

(Lycopodiaceae)

with

biological

activities

incluodmiHnuperzia

serrata

,

are

two

repres

g

AChE

inhibitory

activity，and

separated

into

fouresntatives

of

the

lycodine

class

of

Ltructural

classes:

the

lycopodines,

the

lycodines,

ptohdeium

alkaloids

and

exhibit

potentnhibitory

activity

against

AChE.

6A

has

already

been

marketed

as

a

new

drug

in

the

treatment

of

AD

in

Chin

a.椭圆马尾杉属于石杉科，分布在中国广西省和云南省，以及越南北部。这种植物能生产大量的石杉碱甲，但没有详细的化学成分调查已经完成。本文，我们从马尾杉中分离出五种新的lycodine（石松定）型石松生物碱，phleghenrines

A−D（1-4）和neophleghenrineA（5），以及石杉碱甲和石杉碱乙。

1-5的结构和绝对构型用使用广泛的光谱数据加上计算测定 抗乙酰胆碱酯酶和丁酰胆碱酯酶活性acehin

.ng

tleg

el

cinhibitory

activity

against

AChE

and

BuChE.Aim

Phlegmariurus

henryi

(Baker)

Ching,

the

family

Huperziae

,

distributed

in

the

Guangxi

and

Yunan

provinces

in

Cna,

and

in

northern

Vietnam.

produce

large

amounts

of

HupA

,

but

no

detailed

chemical

constituent

investigatio

the

isolation

of

five

new

Lycopodium

alkaloids

belongio

the

lycodine

class,

phleghenrines

A−D(1−4)

and

neoph

ghenrine

A

(5),

together

with

HupA

(6)

and

HupB

(7)

fromP.

henryi.

The

structuresand

absolute

configurations

of

1−5

usinxtensive

spectroscopic

data

coupled

with

computationaalculation.Phlegmariurus

henryi(Baker)ChingExperimental02植物材料：椭圆马尾杉全草植物，云南省锦屏Extraction

and

IsolationPlant

Material：The

whole

plant

of

P.

henryi

was

collected

from

Jinping,

Yunnan

Province,

People’s

Republic

of

ChinaThe

air-dried

and

powdered

sample

(4.8

kg)

was

extracted

with

pure

methanol

for

three

times

(4,

3,

and3

h),

and

the

extract

was

patitioned

between

EtOAcand

H2O

with

1%

hydrochloric

acid.

The

water-soluble

portion

was

adjusted

to

pH

9

with

saturated

sodium

carbonate

solution,

and

then

was

partitioned

withCHCl3

to

afford

an

alkaloidal

extract

(8.2

g).1.中压柱层析（MCI凝胶柱）甲醇/水(1:9,3:7,6:4,8:2,1:0)；2.硅胶柱氯仿/甲醇（9:1从到1:1）；3.硅胶柱氯仿-甲醇（9:1

4.硅胶柱石油醚、丙酮、二乙胺4:1:1

；5.硅胶柱氯仿/甲醇再乙酸乙酯-甲醇（6:4）

6.羟丙基葡聚糖凝胶柱甲醇；Column

chromatography

(CC)①②③④⑤⑥Column

chromatographymobile

phase①MPLC

(MCI

gel)CH3OH/H2O

(1:9,

3:7,

6:4,

8:2,

1:0)②silica

gel

columnCHCl3/CH3OH

(from

9:1

to

1:1)③silica

gel

columnCHCl3

/CH3OH

(9:1)④silica

gel

CCpetroleum

ether/acetone/diethylamine

(4:1:1)⑤silica

gel

CCCHCl3/CH3OH；EtOAc/CH3OH

(6:4)⑥Sephadex

LH-20

columnmethanol⑧

.⑨

.⑩

.➃

.⑫

.⑬

⑭..⑦

⑮..7.硅胶柱乙酸乙酯/甲醇/二乙胺等度洗脱系统（6:1:2）；8.硅胶柱’氯仿-甲醇梯度洗脱系统（8:1～1:1）；9.羟丙基葡聚糖凝胶柱甲醇；10.硅胶柱乙酸乙酯-甲醇梯度洗脱系统（6:4）；11.硅胶柱乙酸乙酯-甲醇（7:3）；12.硅胶柱氯仿-甲醇（从1

5:1到5：1）13.羟丙基葡聚糖凝胶柱甲醇14.硅胶柱氯仿-甲醇（9:1）；

15.半制备反相高效液相色谱22%甲醇水溶液中含0.1%三氟乙酸Column

chromatographymobile

phase⑦

. silica

gel

CC⑧

. silica

gel

CC⑨

. Sephadex

LH-20

column⑩

. silica

gel

coloumn➃

. silica

gel

CC⑫

. silica

gel

column⑬

. SephadexLH-20⑭

. silica

gel

CC⑮

. semi-preparative

reversed-phaseHPLCEtOAc/CH3OH/diethylamine

(6:1:2)CHCl3/CH3OH

(from

8:1

to

1:1)methanolEtOAc/CH3OH(6:4)EtOAc/CH3OH(7:3)CHCl3/CH3OH(from

15:1

to

5:1)methanolCHCl3/CH3OH(9:1)22%

CH3OH

in

H2O

containing

0.1%trifluoroacetic

acid旋光度，佳司科（Jasco）p-1020旋光仪;紫外光谱，岛津uv-2401a分光光度计甲醇溶液，扫描190-400

nm;电子圆二色谱(ECD)甲醇溶液应用光物理公司Chirascan圆二色光谱仪从195-400

nm扫描;所有氢谱，碳谱和二维NMR谱(HSQC,1H–1HCOSY,HMBC,ROESY)，布鲁克AV

iii-600光谱仪600MHz氢谱和150MHz碳谱质谱，API-Qstar-Pulsar质谱仪(ESI（电喷雾离子源）和HRESI（高分辨率电喷雾离子源）模式)spectroscopic

analysisOptical

rotations:Jasco

P-1020

polarimeterUV

spectra:Shimadzu

UV-2401A

spectrophotometerElectronic

Circular

dichroism

(ECD)

spectra:Applied

Photophysics

ChirascanmeterAll

1

H,

13

C,

and

2D

NMR

(HSQC,

1H–1H

COSY,

HMBC,

ROESY)

spectra:Bruker

AV

III-600

spectrometer

at

600

MHz

for

1

H

and

150

MHz

for

13

C

nucleiMass

spectra

:API-Qstar-Pulsar

instrument

(ESI

and

HRESI

mode)Acetylcholinesterase

(AChE)/Butyrylcholinesterase

Inhibitory

AcResults

and

discussion03namem/z1 Phleghenrine

A256.15622 Phleghenrine

B256.15703 Phleghenrine

C258.17234 Phleghenrine

D250.11135

NeophleghenrineA258.1732molecularformulaC16H20N2OC16H20N2OC16H22N2OC16H14N2OC16H22N2O2和1是非对映异构体a吡啶酮环structures

of

Phleghenrine

A(1)

and

(2)a)

planar

structure:2

and

1

were

diastereomersUV

absorptions

(λ

max

):236

and

315

nmα-pyridone

ring1H

and

13C

NMR

spectrum:一个甲基在δH

1.87(s,H3-16)和三个烯质子在δH

6.00(brd,J=6.6

Hz,H-8),6.32(d,J=9.0

Hz,H-2)，和7.41(d,J=9.0

Hz,H-3);碳谱和DEPT谱（无畸变极化转移增强核磁共振，13C谱图中的伯碳、仲碳、叔碳和季碳）表现出16个碳信号归属于一个甲基，一个α-吡啶基团（δC

165.8，146.0，145.7，117.6，和117.1），一个三取代双键(δ

C142.2

and

125.8)，一个氨基酸次甲基(δ

C

59.7)，三个次甲基，和四个亚甲基。12no.δHδCδHδC1165.8165.826.32,

d

(9.0)117.66.28,

d

(9.0)116.837.41,

d

(9.0)145.77.31,

d

(9.0)146.84117.1118.95146.0145.96a2.83,

dd

(18.6,

3.6)36.52.92,

dd

(18.6,

3.6)31.86b2.76,

dd

(18.6,

3.6)2.65,

dd

(18.6,

3.6)72.50,

m36.22.53,

m35.786.00,

br

d

(6.6)125.85.87,

dt

(6.0,

1.2)126.19a3.23,

m41.42.96,

ddd

(13.2,

10.2,4.8)41.99b3.16,

m2.71,

ddd

(13.2,

9.6,

4.8)10a1.89,

overlapped18.51.80,

m22.310b1.75,

m1.43,

m11a1.89,

overlapped25.11.43,

m22.311b1.37,

m1.34,

m122.43,

m37.51.87,

m37.6133.87,

d

(8.4)59.73.08,

dd

(10.8,

4.8)58.6Table

1.

1

H

(600

MHz)

and

13

C

(150

MHz)

NMR

Data

for1

and

2

in

CD3OD

(δ

in

ppm,

J

in

Hz)d,

J

=

6.6

Hz,

H-8)

;6.32

(d,

J

=

9.0

H1H

spectrum:a

methyl

at

δH

1.87

(s,

H3-16);

three

olefinic

protons

at

δH

6.00

(bH-2);

7.41

(d,

J

=9.0

Hz,

H-3);13C

and

DEPT

NMR

spectra:16

carbon

signalsa

methyl

(δC

22.4)an

α-pyridone

moiety

(δC

165.8,

1460,145.7,

117.6,

and

117.1)a

trisubstituted

double

bond

(δ

C

142

and

125.8)an

aminomethine

(δ

C

59.7)three

methines,

and

four

methylenes结合化学位移（δC

41.4和59.7，分别）和H2-9

与C-13的HMBC谱（远程偶合的碳氢关系）可知C-9和C-13通过一个氮原子连接。此外,H2-9/H2-10/H2-11/H-12/H-13的质子自旋系统可由1H−1H

COSY谱（同核化学位移谱，自选系统中质子之间的关系）看出。哌啶环（B环）2D

NMR

(

1H−

1H

COSY,

HSQC,

and

HMBC)

spectra

:HMBC

correlations

from

H2

-9

to

C-13C-9

and

C-13

(δ

C

41.4

and

59.7,

respectively)1H−

1H

COSY

spectrum:

a

proton

spin

systemof

H2

-9/H2

-10/H2

-11/H-12/H-13HMBC

correlations

of

H2-6

and

H-14

with

C-4

andC-5,

as

well

as

a

proton

spin

system

of

H2

-6/H-7/H-12/H-13/H-14

found

in

1H−

1H

COSY

spectrum.connection

of

C-9

and

C-13

throu

gh

a

nitrogen

atoma

piperidine

ring

(B

ring)A

cycloheptane

ringthe

HMBC

correlations

from

H-14

and

H-7

to

the

doublebond

of

C-8−C-15

in

combination

with

the

proton

spinsystem

of

H-7/H-8

deduced

from

the

1H−

1H

COSY

spectruma

trisubstituted

double

bond

bridge

between

C-7and

C-14the

bicyclo[3.2.2]-nonane

core.ROESY谱（测量氢-氢的距离相关，对于分子量在1000-2000道尔顿的化合物，ROESY比较理想）化合物1和2具有罕见的船式构象zai

b环b)

The

relative

configurations

of

1

and

2by

the

correlations

of

H-13

with

H-9b,

and

H-12

with

H-10b

in

both

cases，compoundsand

2

possess

a

rare

boat

conformation

in

B

ring.通过比较实验和玻尔兹曼加权TD-DFT计算ECD光谱。c)

absolute

configurations

of

1

and

2by

comparison

of

their

experimental

and

Boltzmann-weighted

TD-DFT

calculated

ECD

spectra.(7R,12S,13S,14R)-1

and

(7R,12R,13R,14R)-2.3和1的氢谱和碳谱数据差异在A环。3没有C-2-C-3双键，而是饱和碳δC

31.5

和29.1。C-1(δC

173.3)。3的立体结构被鉴定为7R，12S，13s，14r，和1一样，结合ROESY相关谱，实验电子圆二色谱和计算电子圆二色谱。structures

of

Phleghenrine

C

(3)

and

Neophleghenrine

A

(5)Neophleghenrine

A

(5)

has

an

identical

molecular

formula

with

Phleghenrine

C

(3)1

H

and

13

C

NMR

data

of

3(comparable

with

1):a

marked

difference

in

the

A

ring,no

Signals

for

the

C-2−C-3

double

bond

and

replaced

bytwo

saturated

carbons

at

δ

C

31.5

and

29.1.stereochemistry

of

3:the

ROESY

correlations

of

H-12

with

H2-6

(overlapped

signals),

and

H-12

and

H-13

with

H-10b,

as

well

as

the

comparable

experimental

ECD

spectrum

with

the

calculated

ECD

spectrum(7R,12S,13S,14R)-3与3的碳谱相比，只有三的低信号。C-5-C-4双键δC

175.1

。δ

C

51.6(d,C-4)和78.7(s,C-5)信号被检测出来，结合碳谱和DEPT谱，和HMBC谱。C-5(δC78.7)相对的低场化学位移HMBC相关谱h-9b和C-5之间，以及ROESY相关谱H-6a和H-10a，被个被观察出来(Figure4)，支持′N-2和C-5的联动。因此，5的平面结构包9-含杂氮金刚烷结构。13C

NMR

spectrum

of

5(Compared

with

3):only

three

downfield

signals.

the

C-4−C-5

double

bond

δC

175.1

,δC

51.6(d,

C-4)

and

78.7

(s,

C-5)HMBC

correlation

between

H-9b

and

C-5,together

with

aROESY

correlation

of

H-6a

with

H-10a

,

and

the

relative

downfield

chemical

shift

of

C-5

(δC78.7).the

linkage

of

N-2′

and

C-5Thus,the

planar

structure

of

5

contains

a

9-azaprotoadamantane

moiety.1H−1H

COSY

(bold)

and

selected

HMBC

(arrows)

and

ROESY(double

arrows)

correlations

of5.不饱和度11，这是有报道以来不饱和度最高的石松生物碱。有一个α-吡啶酮，三个低氢信号δH

8.22(dd,J=4.8,1.5

Hz,H-9),7.49(dd,J=7.8,1.5

Hz,H-11),和7.06(dd,J=7.8,4.8

Hz,H-10),和五碳信号δC

163.6(s,C-12),145.9(d,C-9),134.4(s,C-12),132.8(d,C-11),和121.5(d,C-9)。这些表明1的哌啶环（B环）在4中被吡啶环取代。structures

of

Phleghenrine

D

(4)1H−1H

COSY

(bold)

and

selected

HMBC

(arrowcorrelations

of

44

had

11

degrees

of

unsaturation,

which

is

more

than

any

other

reported

Lycopodium

alkaloid

monomer

in

the

literature.1H

and

13C

NMR

spectra(comparing

with

1):the

signals

of

the

α-pyridone

group

and

a

trisubstituted

double

bond

ascribed

to

C-8−C-15,

three

additional

downfield

hydrogen

signals

at

δH

8.22

(dd,

J=

4.8,

1.5

Hz,

H-9),

7.49

(dd,

J

=

7.8,1.5

Hz,

H-11),

and

7.06

(dd,

J=

7.8,

4.8

Hz,

H-10),

and

five

carbon

signals

at

δC

163.6

(s,

C-12),145.9

(d,

C-9),134.4

(s,

C-12),

132.8

(d,

C-11),

and