Tuning Pharmacokinetics to Improve Tumor ... - ACS Publications

Subscriber access provided by University of Sunderland

Article

Tuning Pharmacokinetics to Improve Tumor Accumulation of a PSMA-Targeted Phototheranostic Agent Kara Harmatys, Marta Overchuk, Juan Chen, Lili Ding, Ying Chen, Martin G Pomper, and Gang Zheng Bioconjugate Chem., Just Accepted Manuscript • DOI: 10.1021/acs.bioconjchem.8b00636 • Publication Date (Web): 16 Oct 2018 Downloaded from http://pubs.acs.org on October 17, 2018

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Bioconjugate Chemistry

1

Tuning Pharmacokinetics to Improve Tumor

2

Accumulation of a PSMA-Targeted

3

Phototheranostic Agent

4

Kara M. Harmatys1,4,†, Marta Overchuk1,2,†, Juan Chen1, Lili Ding1, Ying Chen3, Martin G.

5

Pomper3, Gang Zheng*,1-2,4

6 7

†

8

1

9

Toronto, Ontario M5G 1L7, Canada.

Denotes equal contribution Princess Margaret Cancer Centre, University Health Network, 101 College Street,

10

2

11

College Street, Toronto, Ontario M5S 3G9, Canada

12

3

13

States

14

4

15

Ontario M5G 1L7, Canada

16

*

Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164

Johns Hopkins Medical School, 1550 Orleans Street, 492 CRB II, Baltimore, MD 21287, United

Department of Medical Biophysics, University of Toronto, 101 College Street, Toronto,

Corresponding author

17

ACS Paragon Plus Environment

1

Bioconjugate Chemistry 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

18

Abstract

19

We describe a simple and effective bioconjugation strategy to extend the plasma circulation of a

20

low molecular weight targeted phototheranostic agent, which achieves high tumor accumulation

21

(9.74 ± 2.26 %ID/g) and high tumor-to-background ratio (10:1). Long-circulating

22

pyropheophorbide (LC-Pyro) was synthesized with three functional building blocks: 1) a

23

porphyrin photosensitizer for positron-emission tomography (PET)/fluorescence imaging and

24

photodynamic therapy (PDT), 2) a urea-based prostate-specific membrane antigen (PSMA)

25

targeting ligand and 3) a peptide linker to prolong the plasma circulation time. With porphyrin’s

26

copper-64 chelating and optical properties, LC-Pyro demonstrated its dual-modality

27

(fluorescence/PET) imaging potential for selective and quantitative tumor detection in

28

subcutaneous, orthotopic and metastatic murine models. The peptide linker in LC-Pyro

29

prolonged its plasma circulation time about 8.5 times compared to its truncated analog. High

30

tumor accumulation of LC-Pyro enabled potent PDT, which resulted in significantly delayed

31

tumor growth in a subcutaneous xenograft model. This approach can be applied to improve

32

pharmacokinetics of the existing and future targeted PDT agents for enhanced tumor

33

accumulation and efficacy.


Page 2 of 32

2

Page 3 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


34

Background

35

Photodynamic therapy (PDT) has emerged as a highly effective tool for cancer ablation without

36

the use of ionizing radiation and with minimal off-target toxicity.1 The mechanism of PDT relies

37

on the generation of reactive oxygen species (ROS) with a combination of a photosensitizer,

38

light and oxygen.2 Current photosensitizers can be separated as hydrophilic agents that clear

39

rapidly and act intravascularly or hydrophobic agents that accumulate in the tissues.3 Nonspecific

40

uptake of hydrophobic photosensitizers results in off-target light toxicity, which limits the utility

41

of this approach. One way to reduce off-target toxicity and improve photosensitizer tumor

42

accumulation is to design a cancer cell-targeted agent that would confine ROS generation

43

selectively to cancer cells.

44 45

Prostate specific membrane antigen (PSMA) targeting has recently attracted significant attention

46

in the oncology community due to the success of PSMA-targeted nuclear imaging and

47

therapeutic radionuclide delivery, which is beginning to affect management of patients with

48

prostate cancer.4-7 PSMA is a type II transmembrane glycoprotein that is highly overexpressed in

49

prostate cancer. Its expression correlates with cancer aggressiveness.8-10 A variety of PSMA-

50

targeted photosensitizers have appeared recently.7, 11-16 For example, Chen et al. developed a

51

small-molecule PSMA-targeted photosensitizer11, while Nagaya et al. conjugated a near-infrared

52

photosensitizer to a monoclonal antibody.17 While the low-molecular-weight photosensitizer

53

enables deep tumor tissue penetration and fast targeting kinetics, its rapid clearance resulted in a

54

suboptimal efficacy, explaining the need for multiple re-injections followed by PDT. On the

55

contrary, the antibody-based photosensitizer exhibited a long circulation time and favorable

56

biodistribution18, but poor tissue penetration limited its therapeutic potential.19 To address


3


Page 4 of 32

57

limitations of existing PSMA-targeted photosensitizers we designed an agent that combines the

58

virtues of low molecular weight (< 2 kDa) and synthetic accessibility demonstrated by small

59

molecules, while maintaining the long circulation time characteristic of antibody-photosensitizer

60

conjugates.

61 62

The designed agent consists of a pyropheophorbide a photosensitizer, a highly selective PSMA-

63

binding ligand and a peptide-based pharmacokinetic modulator20 (Figure 1). We demonstrate that

64

the insertion of a peptide linker significantly prolongs its plasma circulation time and ultimately

65

enhances its tumor accumulation. This enables efficient single-dose photodynamic treatment,

66

while inherent fluorescence and 64Cu-chelating porphyrin properties allows multimodal imaging

67

(fluorescence/PET) of prostate cancer.

68 69

Figure 1. Schematic structure of the theranostic probe LC-Pyro (long-circulating pyropheophorbide a), which is

70

comprised of three building blocks: 1) A porphyrin photosensitizer capable of deep-red fluorescence imaging and

71

64

72

circulation to promote tumor accumulation; and 3) a high-affinity urea-based small-molecule PSMA targeting

73

ligand.

Cu-chelated PET imaging; 2) a 9-amino acid d-peptide sequence that imparts water-solubility and prolongs plasma

74


4

Page 5 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


75

Results and Discussion

76

LC-Pyro synthesis and characterization

77

We developed a PSMA-targeted phototheranostic agent that consists of three functional building

78

blocks: 1) a porphyrin capable of multimodal fluorescence/PET imaging and photodynamic

79

activity, 2) a 9-amino acid d-peptide linker to impart water-solubility and improve the plasma

80

circulation time and 3) a urea-based high-affinity PSMA as a targeting ligand (Figure 1). LC-

81

Pyro (Long-circulating pyropheophorbide a) and SC-Pyro (Short-circulating pyropheophorbide

82

a) were synthesized and confirmed by UPLC-MS analysis with identified ESI+ mass

83

spectrometry and corresponding UV-vis absorption. (Figure 2A and Figure S1). LC-Pyro (m/z

84

calculated [M]+ 1835.99, found [M]+ 1836.3, [M]2+ 918.0); SC-Pyro (m/z calculated [M]+

85

1119.33, found [M]+ 1119.4, [M]2+ 559.6). DCIBzL was previously synthesized and was used as

86

an inhibitor ligand in vitro and in vivo to confirm PSMA specificity.21 LC-Pyro absorbance

87

(Figure 2B) and fluorescence (Figure 2C) were collected and its measured photodynamic activity

88

revealed an increase in generation of ROS with an increase in laser light dose up to 5 J/cm2

89

(Figure 2D). Next, for PET imaging and quantitative biodistribution studies, we chelated 64Cu to

90

LC-Pyro and evaluated its radiochemical purity by radio-HPLC. The corresponding HPLC traces

91

demonstrated peak alignment of porphyrin absorbance and 64Cu radioactivity, indicating

92

effective 64Cu labeling of LC-Pyro with no remaining free 64Cu.


5


Page 6 of 32

93 94

Figure 2. Structures of PSMA conjugates, the photonic properties of LC-Pyro, and the radio-HPLC trace of 64Cu-

95

LC-Pyro. (A) Structures of LC-Pyro (Long-circulating pyropheophorbide a), SC-Pyro (Short-circulating

96

pyropheophorbide a) and DCIBzL. (B) LC-Pyro absorbance spectrum and (C) fluorescence emission spectrum. (D)

97

Reactive oxygen species generation from LC-Pyro in solution with respect to 671-nm laser light dose (n = 3 from

98

three independent measurements). (E) Evaluation of radiochemical purity of 64Cu-labeled LC-Pyro by radio-HPLC

99

monitored by absorption at 410 nm (left, LC-Pyro signal) and by radioactive detector (64Cu signal). The only peak at

100

retention time of 11 min showed both 410 nm and radioactive signal indicated high purity of 64Cu-labeled LC-Pyro.


6

Page 7 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


101

PSMA affinity ligand demonstrates high targeting specificity

102

Targeted uptake of a photosensitizer by biomarker-overexpressing cancer cells can introduce an

103

additional level of selectivity for PDT. Western blot in Figure 3A validated PSMA expression in

104

the primary or metastatic lines (ML) modified to express high (PSMA+ PC3 PIP; PC3-ML-

105

1124) or low (PSMA- PC3 flu; PC3-ML-1117) levels of PSMA. As cell-penetrating properties of

106

porphyrins22 might contribute to LC-Pyro intracellular uptake in vitro beyond the PSMA-

107

mediated pathway (Figure S2), we synthesized a FITC-labeled analog (LC-FITC) to investigate

108

the targeting selectivity of the peptide-PSMA moiety. Fluorescence microscopy of LC-FITC in

109

Figure 3B revealed selective membrane staining in PC3 prostate cells overexpressing PSMA

110

(PSMA+ PC3 PIP), whereas negligible fluorescence was observed in cells with low PSMA

111

expression (PSMA- PC3 flu) under equivalent incubation settings (3 µM, 3 h) and exposure time.

112

LC-FITC also localized to one focus within the perinuclear region, which has been observed

113

previously and described to represent the mitotic spindle poles or an endosomal compartment.23

114

Addition of 100× excess DCIBzL to LC-FITC achieved successful PSMA binding inhibition,

115

indicating target specificity of the conjugated small-molecule ligand. LC-FITC selectivity was

116

confirmed by flow cytometry over a 22 h. Fluorescence intensity from LC-FITC uptake

117

increased in a time-dependent manner in PSMA+ PC3 PIP cells with a 15-fold higher uptake

118

than PSMA- PC3 flu cells (Figure 3C).


7


Page 8 of 32

119 120

Figure 3. PSMA targeting selectivity and specificity in vitro. (A) Western blot of PC3, PSMA- PC3 flu, PSMA+

121

PC3 PIP, PC3-ML-1117 and PC3-ML-1124 cell lysates using anti-PSMA and β-actin antibodies. (B) Representative

122

fluorescence micrographs of LC-FITC (3 µM, 3 h incubation) uptake in PSMA- PC3 flu and PSMA+ PC3 PIP cells

123

in the presence of 100× excess DCIBzL for target blockade. (C) Representative flow cytometry histogram plots with

124

time-dependent quantification of LC-FITC uptake in PSMA- PC3-flu (open black square) and PSMA+ PC3-PIP

125

(solid green circle) cells. Scale = 25 µm. Each point represents the median ± SD of three independent

126

measurements.

127


8

Page 9 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


128

Peptide linker prolongs plasma circulation time enhancing tumor accumulation of LC-Pyro

129

To validate the pharmacokinetic role of the peptide linker in LC-Pyro and its ability to

130

accumulate in PSMA+ tumors, we synthesized a truncated derivative, SC-Pyro, where the 9-

131

amino acid peptide sequence was replaced by a single lysine residue. Results in Figure 4A

132

demonstrated that LC-Pyro exhibited an 8.5-fold longer plasma circulation time (t1/2, slow = 10.00

133

h; t1/2, fast = 0.50 h) than SC-Pyro (t1/2, slow = 1.17 h; t1/2, slow = 0.20 h). To ensure that the in vivo

134

targeting results were not affected by any difference in binding affinity, the inhibitory activities

135

of LC-Pyro, SC-Pyro and DCIBzL against PSMA were determined using a fluorescence-based

136

assay. All three compounds demonstrated sub-nanomolar inhibitory capacity, despite

137

conjugation of the porphyrin peptide linker to the PSMA inhibitor resulting in an order of

138

magnitude lower PSMA inhibitory properties than DCIBzL (Figure 4B). the specificity of LC-

139

Pyro to PSMA was confirmed in a subcutaneous PSMA+ PC3 PIP xenograft model by inhibition

140

with excess DCIBzL. Excised PSMA+ PC3 PIP tumors revealed less accumulation of LC-Pyro

141

when DCIBzL (PSMA blocker) was injected 30 min beforehand (Figure 4C). Additionally, the

142

PSMA target specificity in vivo was validated with LC-Pyro absent the PSMA affinity ligand.

143

Mice bearing dual subcutaneous PSMA- PC3 flu and PSMA+ PC3 PIP tumors were injected

144

with LC-Pyro (-PSMA affinity ligand) followed by whole-animal in vivo fluorescence imaging

145

(Figure S3). Negligible accumulation of probe within both the PSMA- and PSMA+ tumors was

146

observed after 24 h. Next, to assess the influence of the plasma circulation time between LC-

147

Pyro and SC-Pyro on tumor accumulation and biodistribution, equivalent doses of each agent

148

were intravenously administered to mice bearing dual subcutaneous PSMA- PC3 flu and

149

PSMA+ PC3 PIP tumors followed by whole-animal in vivo fluorescence imaging over 24 h.

150

Figure 4D shows a strong diffuse fluorescence signal from LC-Pyro after 1 h with selective


9


Page 10 of 32

151

accumulation to the PSMA+ after 24 h. There is negligible accumulation within both the PSMA-

152

and PSMA+ tumors 24 h post-administration of SC-Pyro (Figure 4E and Figure S4), indicating

153

the importance of long plasma circulation time for successful tumor accumulation. Ex vivo organ

154

distribution in Figure 4F reveals liver and kidney clearance of LC-Pyro, whereas a majority of

155

SC-Pyro was metabolized and collected in the gallbladder (Figure 4G). That suggests that SC-

156

Pyro cleared rapidly from the body, and therefore, a much higher dose or multiple doses would

157

be required to achieve comparable tumor accumulation to LC-Pyro after 24 h. Ex vivo organ

158

distribution of LC-FITC after 24 h shows similar organ distribution to LC-Pyro, confirming the

159

role of peptide linker in the clearance pathway of the conjugates (Figure S5).

160 161

Figure 4. The pharmacokinetic role of the peptide linker in LC-Pyro for its ability to accumulate in PSMA-

162

expressing tumors. (A) Blood clearance curves of LC-Pyro and SC-Pyro (n = 5 each group) in BALB/c mice with

163

blood samples collected over 48 h. The profiles fit into a two-compartment model with a half-life of 10.00 h for LC-

164

Pyro and 1.17 h for SC-Pyro. (B) Table including the Ki inhibitory activities of SC-Pyro, LC-Pyro and DCIBzL

165

against PSMA determined using a fluorescence-based assay. (C) PSMA inhibition in vivo with 150× molar excess

166

of DCIBzL intravenously injected 30 min prior to LC-Pyro intravenous injection. Mice were sacrificed 24 h post-


10

Page 11 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


167

injection and tumors were excised for ex vivo fluorescence comparison. (D) Representative fluorescence images of

168

mice bearing dual PSMA+ PC3 PIP (red arrow) and PSMA- PC3 flu (green arrow) tumors that were intravenously

169

injected either with LC-Pyro or (E) SC-Pyro at 0.5, 1, 6 and 24 h post-injection. (F) Fluorescence ex vivo organ

170

distribution of mice injected with LC-Pyro or (G) SC-Pyro including PSMA+ PC3 PIP tumor (insets). All images

171

displayed are comparable with the same integration time.

172


11


Page 12 of 32

173

Multimodal imaging of PSMA+ prostate cancer

174

After validating targeting of LC-Pyro in a subcutaneous PSMA+ PC3 PIP tumor model, we

175

explored its theranostic application in two different mouse models bearing either a primary

176

prostate tumor or metastatic prostate cancer. Due to the intrinsic metal-chelation properties of

177

porphyrin ring structures, LC-Pyro was chelated to 64Cu, which allowed for in vivo PET imaging

178

and biodistribution.24 As demonstrated in Figure 5A with PET/CT imaging, 64Cu -LC-Pyro

179

delineated the orthotopic PSMA+ PC3 PIP tumor 17 h post-injection, whereas the PSMA- PC3

180

flu tumor did not have significant uptake. Biodistribution revealed over 4-fold selective

181

accumulation in PSMA+ PC3 PIP tumor [9.74 ± 2.26 percentage injected dose per gram of tissue

182

(%ID/g)] vs. 2.30 ± 0.09 %ID/g in the PSMA- PC3 flu tumor, which showed no distinguishable

183

signal from the surrounding healthy prostate tissue or pelvic organs (Figure 5B). 64Cu -LC-Pyro

184

accumulated greatest in the liver, kidney and feces, indicative of hepatobiliary and renal

185

clearance.

186 187

To investigate further the potential of LC-Pyro to accumulate selectively in PSMA-expressing

188

malignant tissues, we performed fluorescence imaging of micrometastasis. In situ fluorescence

189

images of fluc+/PSMA+ (PC3-ML-1124) and fluc+/PSMA- (PC3-ML-1117) tumors

190

demonstrated selective uptake of LC-Pyro in the PSMA+ tumor nodule (Figure 5C). Those

191

observations aligned with the previous data obtained in subcutaneous xenografts. Following in

192

situ fluorescence imaging, further verification of LC-Pyro accumulation in metastatic nodules

193

and the surrounding tissues was examined by fluorescence microscopy of DAPI-stained tissue

194

sections. Robust porphyrin fluorescence signal in the PSMA+ metastatic nodule microstructure


12

Page 13 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


195

was observed. Notably, the surrounding muscle tissue demonstrated fluorescence near to that of

196

background, further supporting PSMA+ cell selectivity of LC-Pyro.

197 198

Figure 5. 64Cu-LC-Pyro-enabled PET imaging in an orthotopic prostate cancer model and fluorescence detection of

199

PSMA+ micrometastases with LC-Pyro. (A) Sagittal PET/CT images from one mouse bearing either a PSMA+ PC3

200

PIP or PSMA- PC3 flu orthotopic prostate tumor at 3 h and 17 h after intravenous administration of 64Cu -LC-Pyro.

201

(B) 64Cu-labeled LC-Pyro biodistribution in the tumors and the surrounding organs quantified via gamma counting

202

(n = 4 for PSMA+ PC3 PIP; n = 3 for PSMA- PC3 flu, **P < 0.01; **P 1,000 mm3 or started ulcerating, as stated under the animal care institutional

448

guidelines.

449 450

Statistical analysis

451

A two-tailed Student’s t test was used to determine statistical significance. P-values less than

452

0.05 were considered significant.

453

Acknowledgments

454

The authors would like to acknowledge Lili Ding for performing Western blots and assisting


25


Page 26 of 32

455

with flow cytometry measurements; Dr. Il Minn for PSMA binding affinity Ki values

456

measurement; Deborah Scollard, Teesha Komal and Dr. Nicholas Bernards for help with

457

PET/CT image acquisition and reconstruction and Dr. Warren Folz for MR images (STTARR

458

Innovation Centre); Dr. Raymond Reilly, Dr. Zhongli Cai and Dr. Conrad Chan for the

459

assistance with radio-HPLC. This study was funded by the Terry Fox Research Institute

460

(PPG#1075), the Canadian Institute of Health Research (Foundation Grant #154326), the

461

Canadian Cancer Society Research Institute (704718), the Vanier Canada Graduate Scholarship,

462

the Canada Foundation for Innovation, the Joey and Toby Tanenbaum/Brazilian Ball Chair in

463

Prostate Cancer Research, Prostate Cancer Canada, Natural Sciences and Engineering Research

464

Council of Canada, the Princess Margaret Cancer Foundation, and NIH grants CA134675,

465

CA184228, CA202199 and EB024495.

466


26

Page 27 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


467

Abbreviations

468

LC-Pyro, Long-circulating pyropheophorbide a; prostate-specific membrane antigen; PET,

469

positron emission tomography; PDT, photodynamic therapy; kDa, kilodaltons; ROS, reactive

470

oxygen species; SC-Pyro, short-circulating pyropheophorbide a; uPLC-MS, ultra-performance

471

liquid chromatography mass spectrometry; ESI, electrospray ionization; HPLC, high-

472

performance liquid chromatography; J/cm2, Joules per square centimeter; ML, metastatic line;

473

PC3, prostate cancer cell line; LC-FITC, long-circulating fluorescein isothiocyanate; SD,

474

standard deviation; t1/2, half-life; Ki, inhibitor constant; PET/CT, positron emission

475

tomography/computed tomography; %ID/g, percent injected dose per gram; fluc, firefly

476

luciferase ; DAPI, 4′,6-diamidino-2-phenylindole; H&E, Hematoxylin & Eosin; TUNEL,

477

Terminal deoxynucleotidyl transferase dUTP nick end labeling; HBTU, (benzotriazol-1-yl)-

478

N,N,N′,N′-tetramethyluronium hexafluorophosphate; Fmoc, Fluorenylmethyloxycarbonyl

479

protecting group; NHS, N-hydroxysuccinamide; FBS, fetal bovine serum; EDTA,

480

ethylenediaminetetraacetic acid; DMSO, dimethyl sulfoxide; FACS, Fluorescence-activated cell

481

sorting; PBS, phosphate buffered saline; MRI, magnetic resonance imaging; OCT, optical cutting

482

temperature; mg/kg, milligrams per kilogram; LP, longpass; rpm, revolutions per minute; mCi,

483

millicuries; Cy5, cyanine 5.

484

Associated Content

485

The Supporting Information is attached.

486

Flow cytometry, fluorescence mouse imaging and biodistribution, 64Cu-LC-Pyro PET/CT

487

imaging and organ gamma counting, PDT histological organ toxicity, blood clearance of YC-9.

488


27


489

Author Information

490

Corresponding Author

491

*E-mail: [email protected]

492

Notes

493

The authors declare no conflict of interest.

494

Author Contributions

495

Kara M. Harmatys and Marta Overchuk have contributed equally to the manuscript.

Page 28 of 32

496


28

Page 29 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


497

References

498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540

1. Cengel, K. A. S. C. B.; Glatstein, E., PDT: What's Past Is Prologue. Cancer Research 2016, 76 (9), 2497-2500. 2. Wilson, B. C.; Patterson, M. S., The physics, biophysics and technology of photodynamic therapy. Physics in medicine and biology 2008, 53 (9), R61-R109. 3. Ethirajan, M.; Chen, Y.; Joshi, P.; Pandey, R. K., The role of porphyrin chemistry in tumor imaging and photodynamic therapy. Chem Soc Rev 2011, 40 (1), 340-62. 4. Haberkorn, U.; Eder, M.; Kopka, K.; Babich, J. W.; Eisenhut, M., New Strategies in Prostate Cancer: Prostate- Specific Membrane Antigen (PSMA) Ligands for Diagnosis and Therapy. Clinical Cancer Research 2016, 22 (1), 9-15. 5. Kratochwil, C.; Bruchertseifer, F.; Giesel, F. L.; Weis, M.; Verburg, F. A.; Mottaghy, F.; Kopka, K.; Apostolidis, C.; Haberkorn, U.; Morgenstern, A., 225Ac-PSMA-617 for PSMATargeted a-Radiation Therapy of Metastatic Castration-Resistant Prostate Cancer. J Nucl Med 2017, 57, 1941-1945. 6. Kratochwil, C.; Giesel, F. L.; Stefanova, M.; Beneˇ, M.; Bronzel, M.; Afshar-oromieh, A.; Mier, W.; Eder, M.; Kopka, K.; Haberkorn, U., PSMA-Targeted Radionuclide Therapy of Metastatic Castration-Resistant Prostate Cancer with 177Lu-Labeled PSMA-617. J Nucl Med 2017, 57 (8), 1170-1177. 7. Wang, X.; Tsui, B.; Ramamurthy, G.; Zhang, P.; Meyers, J.; Kenney, M. E.; Kiechle, J.; Ponsky, L.; Basilion, J. P., Theranostic Agents for Photodynamic Therapy of Prostate Cancer by Targeting Prostate-Specific Membrane Antigen. Molecular Cancer Therapeutics 2016, 15 (8), 1834-1844. 8. Bostwick, D. G.; Pacelli, A.; Blute, M.; Roche, P.; Ph, D.; Murphy, G. P., Prostate Specific Membrane Antigen Expression in Prostatic Intraepithelial Neoplasia and Adenocarcinoma A Study of 184 Cases. 1998, 2256-2261. 9. Kiess, A. P.; Minn, I.; Vaidyanathan, G.; Hobbs, R. F.; Josefsson, A.; Shen, C.; Brummet, M.; Chen, Y.; Choi, J.; Koumarianou, E.; Baidoo, K.; Brechbiel, M. W.; Mease, R. C.; Sgouros, G.; Zalutsky, M. R.; Pomper, M. G., (2S)-2-(3-(1-Carboxy-5-(4-211AtAstatobenzamido)Pentyl)Ureido)-Pentanedioic Acid for PSMA-Targeted -Particle Radiopharmaceutical Therapy. J Nucl Med 2016, 57 (10), 1569-1575. 10. Israeli, R. S.; Powell, C. T.; Corr, J. G.; Fair, W. R.; Heston, W. D. W., Expression of the Prostate-specific Membrane Antigen. Cancer Research 1994, 54, 1807-1811. 11. Chen, Y.; Chatterjee, S.; Lisok, A.; Minn, I.; Pullambhatla, M.; Wharram, B.; Wang, Y.; Jin, J.; Bhujwalla, Z. M.; Nimmagadda, S.; Mease, R. C.; Pomper, M. G., A PSMA-targeted theranostic agent for photodynamic therapy. Journal of Photochemistry and Photobiology B: Biology 2017, 167, 111-116. 12. Fakhrejahani, F.; Madan, R. A.; Dahut, W. L., Management Options for Biochemically Recurrent Prostate Cancer. Curr Treat Options Oncol 2017, 18 (5), 26. 13. Liu, T.; Wu, L. Y.; Berkman, C. E., Prostate-specific membrane antigen-targeted photodynamic therapy induces rapid cytoskeletal disruption. Cancer Letters 2010, 296 (1), 106112. 14. Liu, T.; Wu, L. Y.; Choi, J. K.; Berkman, C. E., In vitro targeted photodynamic therapy with a pyropheophorbide-a conjugated inhibitor of prostate-specific membrane antigen. The Prostate 2009, 69, 585-594.


29


541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585

Page 30 of 32

15. Nagaya, T.; Nakamura, Y.; Okuyama, S.; Ogata, F.; Maruoka, Y.; Choyke, P. L.; Kobayashi, H., Near-Infrared Photoimmunotherapy Targeting Prostate Cancer with ProstateSpecific Membrane Antigen (PSMA) Antibody. Molecular Cancer Research 2017, 15 (9), 11531162. 16. Serrell, E. C.; A, B.; Pitts, D.; D, M.; Hayn, M.; D, M.; Beaule, L.; D, M.; Hansen, M. H.; D, M.; Sammon, J. D.; O, D., Review of the comparative effectiveness of radical prostatectomy , radiation therapy , or expectant management of localized prostate cancer in registry data. Urologic Oncology: Seminars and Original Investigations 2017, 1-10. 17. Nakajima, T.; Mitsunga, M.; Bander, N.; Heston, W. D.; Choyke, P. L.; Kobayashi, H., Targeted, Activatable, In Vivo Fluorescence Imaging of Prostate-specific Membrane Antigen (PSMA)-positive Tumors Using the Quenched Humanized J591 Antibody-ICG Conjugate. Bioconjugate chemistry 2012, 22 (8), 1700-1705. 18. Watanabe, R.; Hanaoka, H.; Sato, K.; Nagaya, T.; Harada, T.; Mitsunaga, M., Photoimmunotherapy Targeting Prostate-Specific Membrane Antigen: Are Antibody Fragments as Effective as Antibodies? J Nucl Med 2018, 56 (1), 140-145. 19. Minchinton, A. I.; Tannock, I. F., Drug penetration in solid tumours. Nat Rev Cancer 2006, 6 (8), 583-92. 20. Stefflova, K.; Li, H.; Chen, J.; Zheng, G., Peptide-based pharmacomodulation of a cancer-targeted optical imaging and photodynamic therapy agent. Bioconjug Chem 2007, 18 (2), 379-88. 21. Chandran, S. S.; Banerjee, S. R.; Mease, R. C.; Pomper, M. G.; Denmeade, S. R., Characterization of a targeted nanoparticle functionalized with a urea-based inhibitor of prostatespecific membrane antigen (PSMA). Cancer Biol Ther 2008, 7 (6), 974-82. 22. Chen, Y.; Gryshuk, A.; Achilefu, S.; Ohulchansky, T.; Potter, W.; Zhong, T.; Morgan, J.; Chance, B.; Prasad, P. N.; Henderson, B. W.; Oseroff, A.; Pandey, R. K., A Novel Approach to a Bifunctional Photosensitizer for Tumor Imaging and Phototherapy. Bioconjugate Chem 2005, 1264-1274. 23. Kiess, A. P.; Minn, I.; Chen, Y.; Hobbs, R.; Sgouros, G.; Ronnie, C.; Pullambhatla, M.; Shen, C. J.; Foss, C. A.; Martin, G., Auger Radiopharmaceutical Therapy Targeting ProstateSpecific Membrane Antigen. J Nucl Med 2015, 56 (9), 1401-1407. 24. Shi, J.; Liu, T. W.; Chen, J.; Green, D.; Jaffray, D.; Wilson, B. C.; Wang, F.; Zheng, G., Transforming a Targeted Porphyrin Theranostic Agent into a PET Imaging Probe for Cancer. Theranostics 2011, 1, 363-70. 25. Zheng, G.; Li, H.; Zhang, M.; Lund-Katz, S.; Chance, B.; Glickson, J. D., Low-density lipoprotein reconstituted by pyropheophorbide cholesteryl oleate as target-specific photosensitizer. Bioconjug Chem 2002, 13 (3), 392-6. 26. Chandran, S. S.; Banerjee, S. R.; Mease, R. C.; Pomper, M. G.; Denmeade, S. R., Characterization of a targeted nanoparticle functionalized with a urea-based inhibitor of prostatespecific membrane antigen (PSMA). Cancer Biology and Therapy 2008, 7 (6), 974-982. 27. Chen, Y.; Dhara, S.; Banerjee, S. R.; Byun, Y.; Pullambhatla, M.; Mease, R. C.; Pomper, M. G., A low molecular weight PSMA-based fluorescent imaging agent for cancer. Biochem Biophys Res Commun 2009, 390 (3), 624-9. 28. Cheng, Y.; Prusoff, W. H., Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 1973, 22 (23), 3099-108.


30

Page 31 of 32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

586 587 588 589 590 591 592 593 594 595 596 597 598


29. Wilson, B. C.; Firnau, G.; Jeeves, W. P.; Brown, K. L.; Burns-McCormick, D. M., Chromatographic analysis and tissue distribution of radiocopper-labelled haematoporphyrin derivatives. Lasers in Medical Science 1988, 3 (1), 71-80. 30. Chang, S. S.; Reuter, V. E.; Heston, W. D. W.; Bander, N. H.; Grauer, L. S.; Gaudin, P. B., Five Different Anti-Prostate-specific Membrane Antigen ( PSMA ) Antibodies Confirm PSMA Expression in Tumor-associated Neovasculature 1. Cancer Res 1999, (212), 3192-3198. 31. Jin, C. S.; Overchuk, M.; Cui, L.; Wilson, B. C.; Bristow, R. G.; Chen, J.; Zheng, G., Nanoparticle-Enabled Selective Destruction of Prostate Tumor Using MRI-Guided Focal Photothermal Therapy. Prostate 2016, 76 (13), 1169-81. 32. Castanares, M. A.; Copeland, B. T.; Chowdhury, W. H.; Liu, M. M.; Rodriguez, R.; Pomper, M. G.; Lupold, S. E.; Foss, C. A., Characterization of a novel metastatic prostate cancer cell line of LNCaP origin. The Prostate 2016, 76 (2), 215-25.


31


152x62mm (300 x 300 DPI)


Page 32 of 32

Tuning Pharmacokinetics to Improve Tumor ... - ACS Publications

Recommend Documents