Aminatophosphorous(1+) Salts: A New Class of Nonmobile Protectant

Chapter 29

Aminatophosphorous(1+) Salts: A New Class of Nonmobile Protectant Fungicides

Downloaded by PENNSYLVANIA STATE UNIV on June 18, 2012 | http://pubs.acs.org Publication Date: May 14, 1998 | doi: 10.1021/bk-1998-0686.ch029

1

1

1

1

1

John Miesel , CarlDenny ,LeahReimer ,Greg Kemmitt , Todd Mathieson , Mary Barkham , and Sarah Cranstone 2

2

1

Discovery Research Center, DowElanco, 9330 Zionsville Road, Indianapolis,IN46268-1053

2

Letcombe Laboratory, DowElanco Europe, Letcombe Regis, Oxon0W129JT, United Kingdom It was found that members of a series of aryl and heterocyclic aminatophosphorus(1+)salts form a new class of non-mobile protectant fungicides especially efficacious against Plasmopara viticola, the causative organism of grape downy mildew. The synthesis and structure-activity relationships of this class of fungicides are discussed. Development of this series was discontinued due to phytotoxicity to vines and extreme ocular irritation found in the most active compounds.

Non-mobile protectant fungicides such as the dithiocarbamates have a long history of use as mixing partners for crop disease management (7). More recently the use of existing products, mancozeb for example, has come under scrutiny because of their very high rate of application and possible toxicological problems. Newer compounds are needed to fill this niche in the marketplace. Aminatophosphorus( 1 +) salts are a new class of non-mobile protectant fungicides which were examined at DowElanco. Members of this series possess efficacy against several fungal organisms of commercial interest, Plasmopara viticola, Erysiphe graminis, and Puccinia recondata. The activity against P. viticola, the causative agent of grape downy mildew, was the most significant and used to direct the study of the SAR of this series. Plasmopara Screening Methods Key points in the screening methods for P. viticola are outlined below. To be passed from initial to follow-up screening generally required a disease control rating of 7 to 9 at 100 ppm in the initial screening. ©1998 American Chemical Society

In Synthesis and Chemistry of Agrochemicals V; Baker, D., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 1998.

295

296


Initial P. viticola Screening: • 4 week old grape seedlings - single replicate, 2 plants. • Technical material applied at 400, 100, 25, 6.25 ppm. • Inoculation 2 hrs (or 24 hrs) after application. • Read test 4-5 days after inoculation. • Disease control expressed on a modified Barratt-Horsfall scale (2), 9 = 100% control, 8 = 99-97%, 7 = 97-90%, 5 = 74-60%, 1 =0-19%. Follow-up P. viticola Screening: • 4 week old grape seedlings - 3 replicates, 1 plant each. • Technical material applied at 100, 25, and 6.25 ppm. • Inoculation 24, 96, and 240 hrs after application. • Disease control expressed as a percentage relative to untreated control. Compound Structure and Synthesis Aminatophosphorus( 1+) salts are essentially N-alkylated phosphine imines. However the products are best described by the phosphorus to nitrogen single bond structures shown in Figure 1. This structure was confirmed by single crystal X-ray analysis of two members of the series. X

+

Ar-N-PAr R'

3

B—A / C ι R N

+ Hetero-N-PAr

χ

~N—PAr,

3

R' Figure 1 - Structure of Aminatophosphorus(l+) Salts The phosphine imine intermediate is prepared in one of two ways as shown in Figure 2. The Staudinger reaction of an azide with a phosphine is most useful when examining a variety of phosphines but requires the synthesis of sometimes unstable azides (J). The other general synthesis uses in situ generation of a phosphine dibromide followed by reaction with an aniline or aminoheterocycle (4). This route is favored when all imine substituents are aromatic in nature. Alkylation of phosphine imines has been long known (5). The rate of alkylation of the phosphine imine is strongly dependent on its electronegativity, the ease of displacement of the leaving group, and steric factors. When the alkylation solvent is dichloromethane, pure product can be isolated by precipitation on addition of ether or a mixture of ether and ethyl acetate.


297

PhN

S

+ RRR'P

3

° ^

Ar'NH, 2

Ar,PBr,

1

»

Ph^PIH^"

PhH, reflux

+

>

Ar'tt=PAr

Et N PhH, reflux*

3

3

H

e

t

e


Ar 'N=PAr

r

o

N

33

2

H

Solvent, reflux

»

HeteroN=PAr

3

Ar-N—PAr, 1

R

3

x"

Figure 2 - Synthesis of Aminatophosphorus(l+) Salts Structure-Activity Relationships The compounds which are discussed below are only a portion of those synthesized and tested. They are representative of the series and accurately reflect the trends discussed. In some cases, the counter ions will vary in a particular series. We have found no significant difference in fungicidal efficacy due to the counter ion and will not present this data due to space limitations. Effect of Phosphine Substitution on P. viticola Activity. As indicated in Table I, aryl substitution on phosphorus provided higher activity, especially beyond the 2 hour test. Table I - Effect of Phosphine Substituent on P. viticola Activity 4~V-N-PRR'R CH 3

M

r

Activity R Ph Cyclohexyl Ph

Κ Ph Cyclohexyl Ph

Κ Ph Cyclohexyl Bu

4-CH3Ph

4-CH3Ph

4-CH3Ph

2-CH3Ph

2-CH3Ph

2-CH3PI1

4-CH3OPI1

4-CH3QPh

4-CH30Ph

2Hr 9, 6, 1, 1 1 8, 8, 6, 1 9, 9, 9, 3 9, 8, 8, 5 9, 8, 6, 6

24 Hr 53,43, 5 100, 60, 22 97, 50, 6 98, 90, 73

Effect of N-Aryl Substitution on P. viticola Activity. The effect of N-aryl monosubstitution is given below in Table II. Monosubstitution was generally disappointing although compounds with electron-donating substituents were more active than those with electron-withdrawing substituents. The more lipophilic phenoxy group was an improvement especially in the ortho position.


298


Table II - Effect of Aryl Mono-Substituent on P. viticola Activity

Activity 2Hr 24 Hr 9, 6,1,1 9,7, 1, 1 9, 9, 9, 6 100, 93, 87 93, 68,44 9, 9, 9, 9 9, 7, 6, 6 84, 73, 8 28, 10, 5 9, 9, 1, 1

R H 2-OCH3 2-OPh 4-OPh 3-OPh 4-N02

Activity 2Hr 5,1,1,1 7, 3,1,1 9, 6, 6, 2 8, 3,1,1 1, 1, 1, 1 1,1,1,1

R 4-CN 3-CF3 2-C2H5 2-SCH3 2-SOCH3 2-SO2CH3

However, the effect of multiple substitution with electron-donating substituents, especially with one of the substituents in the ortho position, was quite favorable as shown in Table III. Both the 2,4-dimethoxy analog and the dibenzofuran compound 1 provide substantial protection against P. viticola when grape seedlings were infected 10 days after treatment. Table III - Effect of Aryl Substituent on P. viticola Activity

R

T5^

P h

CH R 2-OCH3 2-OCH3 3-OCH3 3-OCH3 2-CH3

K 4-OCH3 5-OCH3 4-OCH3 5-OCH3 4-CH3 1

3

3 1

r

2Hr 9, 9, 8, 6 9, 9, 9, 9 9, 1, 1, 1 8,1,1,1 8, 8, 1, 1 9, 9, 5, 5

Γ Activity 96 Hr 100, 93, 13 92, 67, 20

240 Hr 96, 87, 23

93, 89, 50

87, 77, 57

Effect of N-alkyl substitution on P. viticola activity. The effects of the N-alkyl substituent on P. viticola activity are outlined in Table IV. Simple alkyl groups larger than methyl offered no activity advantage and the more electron-poor cyanomethyl group decreased activity. Benzyl substitution most often improved activity, especially residual activity, but the optimum benzyl substituent would vary with substitution on the N-aryl ring. Throughout the entire series, the in vivo activity of any specific compound appeared to depend on whole molecule properties which could not be predicted by simply combining the individual substituents that displayed the most activity.


299 Table IV - Effect of Alkyl Substituent on P. viticola Activity PPh, R R CH3


Activity 96 Hr

24 Hr 9, 6, 1, 1 9, 7, 6, 1 9, 9, 3, 1 9, 9, 9, 9 9, 9, 8, 8 9, 8, 3, 1

n-Bu i-Bu PhCH2 4-CH30PhCH2 3,4-Cl2PhCH2

240 Hr

100, 85,61

99, 92, 73 69, 0, 0 62,31,5

OMe MeO-4

PPh R

R

X~ Activity 96 Hr 100, 93, 13

24 Hr 9, 9, 8, 6 9, 6,1,1 9, 8, 3, 1 9, 9, 7, 3 9, 9, 7, 3 9, 8, 8, 6

CH3

NCCH2 CH2=CHCH2 PhCH2 4-CH30PhCH2 3,4-Cl2PhCH2

3

240 Hr 96, 87, 23 93,71,61

100,61,2 98, 93, 40 93, 87, 9

100, 99, 82 99, 90, 42

Effect of N-heterocyclic substitution on P. viticola Activity. Synthesis of Nheterocyclic aminatophosphorus( 1 +) salts provides an additional complication. Alkylation can occur endo at the ring nitrogen, as in the thiazole below, or at the exocyclic nitrogen as in the very similar isoxazole. Endo addition occurs in about 80 - 90% of the examples, and mixtures are obtained in a few cases. Proton NMR is diagnostic for the position of alkylation since groups adding exo show vicinal coupling between the α protons on the alkyl group and phosphorous. Those adding endo have the α protons unsplit. Ph CH2CI2,

EndQ Alkylation S

N=PPh,

CH3I

V v

reflux

Ç

Exo Alkylation

N=PPh,

H

3

N-PPh,

CH2CI2, CH^I

reflux

fPh,

Ph

.Ν Ο

I


300 The electronegativity of the N-heterocyclic substituent has a strong effect on biological efficacy. The relationship of ring electron density with activity mirrors that seen with the N-aryl members of the series. The more electron-rich the ring, the greater is the P. viticola control. This is shown in Table V for heterocycles with either phenyl or ί-butyl substituents.


Table V - Effect of Heterocycle on P. viticola Activity Hef=N-ÊPh CH, 3 endo

3

or

A

Hetero

Ph-/

Het-N-fPh, CH, exo

Position

24 Hr

Activity 96 Hr

240 Hr

endo

9, 9,9, 6

86, 67, 33

85, 25, 20

endo

9, 9, 9, 9

98, 97, 77

93, 93, 73

Ν Ν

exo Ph-ί^Ν endo

6, 5, 3, 1

endo

9, 9, 5, 1

endo

9, 9, 9, 7

N-N

endo

9,7,1,1

N=N rBu—(\ /) N-N

endo

6,1,1,1

Ph V Ph

N

N

N-N 96,43,43

The same trend is obtained by varying the electronegativity of a substituent on the heterocyclic ring. This is exemplified in Table VI by a group of pyridine


301 compounds. The benzosubstituent, in this case the quinoline ring, is often an analog equivalent to a phenyl substituent. Table VI - Effect of Heterocycle Substituent on P. viticola Activity Het=N-£ph CH j— 3

Het-N-ÊPh

o r

endo Downloaded by PENNSYLVANIA STATE UNIV on June 18, 2012 | http://pubs.acs.org Publication Date: May 14, 1998 | doi: 10.1021/bk-1998-0686.ch029

3

j~

3

exo Activity

Hetero

N

C

^ N

Position

24 Hr

endo

8, 7, 2, 1

endo

9,3,1,1

endo

3, 1, 1, 1

endo

9, 7, 4, 3

96 Hr

240 Hr

N"

H

H

3°

"N

ι

C

3 v^N

ÇH

3

e n d o

8, 6, 2, 1

exo

9,9,9,6

99,80, 19

endo

9,9,9,9

100,85,70

11 80,52,3

The non-predictability of the effect of the N-alkyl substituent on efficacy, especially residual activity, is demonstrated in Table VII. The thiazole compound 2 has excellent 240 hour residual activity with the N-methyl substituent, but much poorer with the N-benzyl. The reverse is true with the quinoline analogs 3.


302 Table VII - Effect of Alkyl Substituent on P. viticola Activity


2 Compound 2 2 3 3

R CH3 PhCH2 CH3 PhCH2

3 24 Hr 9, 9, 9, 9 8, 7, 1, 1 9, 9, 9, 9 9, 9, 9, 9

Activity 96 Hr 98, 97, 77 100, 99, 68 100, 85, 70 99, 99, 90

240 Hr 93, 93, 73 25, 16, 0 80, 52, 3 97, 97, 8

Pot Trials At DowElanco the next stage in structure-activity relationship testing for P. viticola is outdoor pot tests. Pot tests allow us to examine some of the features of a field test using less compound. Large vines that are initially 2.5 - 3 feet tall and that are hardened to the outdoors are used. The test compound in a more commercial formulation is examined after multiple applications at 7 day intervals. Compounds 1, 4, and 5 were tested in this fashion.

Initial results were disappointing. All three salts were superior to Dithane in the greenhouse. However, in the pot test only compound 5 had an LC90 below Dithane. More troubling was the unexpected appearance of phytotoxicity in the test. The more efficacious 5 was also more phytotoxic. This phytotoxicity was quite characteristic at low rates where leaf cupping and a leathery surface on the affected leaves were observed. This damage progressed to partial leaf necrosis at higher rates. The affected leaves were newly emerged at the time of compound application and the damage appeared as the leaves expanded. Mature leaves were unaffected by compounds in this series. Special Phytotoxicity Testing To continue our examination of this series, it was necessary to reproduce this phytotoxicity in a greenhouse test. Some simple changes in the screen were sufficient. The presence of newly emerged leaves was ensured by using larger vines. The plants were held at 100% humidity for 24 hours after compound application


303 which markedly increased phytotoxicity. Symptom development was encouraged by holding the plants for 10 days. Damage similar to that in the pot test was expressed using this protocol. Based on the data from the special phytotoxicity test for the three pot trial compounds as well as the two other active analogs shown in Table VIII, compound 7 was an obvious choice for further examination.


Table VIII - Special Greenhouse Vine Phytotoxicity Test

Compound 1 4 5 6 7

Percent total leaf area expressing symptoms Rate (ppm) 100 500 1000 5 0 1 13 22 27 15 43 10 15 13 0 0 1.7

0

2

CH (4-MeOPh) 2

6

7

Further examination of compound 7 The pot trial of compound 7 was quite favorable. The efficacy of this compound against P. viticola was equivalent to Dithane at 0.25 the rate. As predicted by the special greenhouse testing, there was no phytotoxicity to vines after 4 applications of 7 at 1000 ppm. Before going to a full field trial, a mammalian hazard evaluation on 7 was conducted. Although non-toxic by oral or dermal routes, the compound was found to be an extremely dangerous ocular irritant. The ocular damage was so severe that the animals were euthanized after 24 hours. No further work with 7 was conducted. Caution: All aminatophosphorus( 1 +) salts should be considered as possible severe occular irritants. Full eye protection should be used when preparing or using compounds in this class of chemistry.


304 Summary


A series of aminatophosphorus(l+) salts was examined as non-mobile protectant fungicides for the control of P. viticola. Structure-activity relationship studies were successful in developing a number of compounds with excellent residual control in the greenhouse. Special trials indicated that many of the most active compounds caused unacceptable phytotoxicity to vines with repeat applications at efficacious rates. Extreme ocular irritation found with the most promising compound in this series ended further work on the series. Acknowledgments The authors would like to thank the other scientists in DowElanco Discovery Research who provided helpful assistance through the course of these studies. Special acknowledgment must be given to Prof. Phil Fanwick of Purdue University for determining the X-ray structures mentioned in this report. Literature Cited 1. Encyclopedia of Chemical Technology, Grayson,M.,Eckworth,D.Eds, John Wiley and Sons: New York, NY, 1980; Vol.11,pp 490-98. 2. Horsfall,J.G.;Barratt,R.W.;Phytopath., 1945, 35, 655. 3. Staudinger,H.;Meyer,J.;Helv. Chim. Acta, 1919, 2, 635. 4. Horner,L.;Oediger,H.;Liebigs Ann., 1959, 627, 142. 5. Staudinger,H.;Hauser,E.;Helv. Chim. Acta, 1921, 4, 861.


Aminatophosphorous(1+) Salts: A New Class of Nonmobile Protectant

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