AU2019400110A1

AU2019400110A1 - Radioactive fluorine-labeled Larotrectinib compound and preparation method therefor

Info

Publication number: AU2019400110A1
Application number: AU2019400110A
Authority: AU
Inventors: Gang Huang; Bin Li
Original assignee: Shanghai University of Medicine and Health Sciences
Current assignee: Shanghai University of Medicine and Health Sciences
Priority date: 2018-12-14
Filing date: 2019-09-19
Publication date: 2021-07-01
Anticipated expiration: 2039-09-19
Also published as: WO2020119205A1; AU2019400110B2; CN109705124B; CN109705124A

Abstract

A radioactive fluorine-labeled Larotrectinib compound and a preparation method therefor, particularly, an in-vivo imaging agent of a Trk receptor subtype in a refractory solid tumor. Provided are a radioactive fluorine compound

Description

RADIOACTIVE FLUORINE-LABELED LAROTRECTINIB COMPOUND AND PREPARATION METHOD THEREOF BACKGROUND OF THE INVENTION

[0001] 1. Technical Field

[0002] The invention relates to the field of chemical drug synthesis, especially to

an in vivo imaging agent for Trk receptor subtype in refractory solid tumors, and more

particularly to a radioactive fluoride 1 8 F-Larotrectinib based on a novel tyrosine receptor

kinase (TRK) inhibitor Larotrectinib and its 18F-Larotrectinib analogs, and more

particularly to a preparation method of the radioactive 18F-labeled compound

18F-Larotrectinib and 18F-Larotrectinib analogs based on Larotrectinib and its analogs.

[0003] 2. Description of Related Art

[0004] Larotrectinib was developed by Loxo Oncology. As a broad-spectrum

tumor drug, it is used for all tumor patients that express (TRK). This small-molecule TRK

inhibitor is highly selective for TRK. By inhibiting the TRK signaling pathway,

larotrectinib can inhibit tumor growth. Larotrectinib is a potent oral TRK inhibitor with

consistent and long-lasting anti-tumor activity in TRK fusion tumors. It is applicable to a

wide range of patient ages and tumor types and its indications are distributed in 13

different tumor types.Larotrectinib has good tolerance, is effective against a variety of

solid tumors in adults and children, including salivary, infantile fibrosarcoma, lung cancer,

thyroid cancer, colon cancer, melanoma (melanoma), cholangio cancer, GISTs, breast

cancer, and various sarcoma cancers. The US FDA (http://www.chemdrug.com/article/11/)

has granted orphan drug designation and breakthrough drug designation to larotrectinib.c

Larotrectinib is expected to become the first therapeutic drug to be developed and

approved simultaneously for adults and children, and it is the first tumor-targeted

therapeutic drug that spans all traditionally defined tumor types and molecular meanings.

The structure of Larotrectinib is as follows:

F

OH N F~Q HN Larotrectinib 0

[0005] However, at present, for the distribution of drug molecules in the body and

the evaluation of curative effect, the positron-emitting radiopharmaceutical

18F-deoxyglucose (18F-FDG) is usually used to indirectly evaluate the curative effect of

the drug on tumors and related diseases. Moreover, the positron-emitting

radiopharmaceutical 18F-deoxyglucose (18F-FDG) also has high FDG absorption in

non-tumor tissues and inflammatory cell components, which may cause false positive

results in tumor diagnosis due to inflammation when FDG is used in tumor imaging.

Therefore, how to on line trace the distribution of Larotrectinib in humans or animals and

the status in solid tumors in vivo, intuitively determine the physiological functions of

Larotrectinib in tumors, and evaluate its curative effect and its healing effect is a pressing

problem.

[0006] PET (Positron emission tomography) is a non-immersive imaging

technology that can realize the visualization and quantitative evaluation of the

physiological and biochemical functions of drug molecules and the pharmacological

process at the molecular level in vivo. As a small molecule inhibitor for tropomyosin

receptor kinase (TRK), Larotrectinib molecule is highly selective for TRK and can be used

as a PET imaging molecule to accurately locate Larotrectinib molecules in vivo to

evaluate tumor conditions, realize online tracing of Larotrectinib molecules in animals or

humans and the status in solid tumors, intuitively determine the physiological function and

pharmacological process of Larotrectinib in animals, humans and tumors, and evaluate its

curative effect and its healing effect. There is no more effective technical means at present.

BRIEF SUMMARY OF THE INVENTION

[0007] The objective of the invention is to provide a 1 [ 8F]-labeled Larotrectinib

compound and a preparation method thereof in order to overcome the above-mentioned

defects in the prior art.

[0008] The objective of the invention can be reached by the following technical

solutions: A [ 1 8F]-labeled Larotrectinib compound, including a [ 18F]-Larotrectinib

compound having the following structural formula and its analogs:

F 18F

_NOH _NN OH

1F (R) N F (R)

18F 0 N47(N HNRN F Ni HN-N 1 0 2 O 18F-2-Larotrectinib 18 F-5-Larotrectinib

18F-2-Larotrectinib 18F-5-Larotrectinib

18F

18F N OH R1(2) N -H

18F N' N NNNb N b HN-If N6HNy

3 0 4 18 18 F-2-Larotrectinib analogues F-5-Larotrectinib analogues

18F-2-Larotrectinib analogues 18F-5-Larotrectinib analogues

[0009] A preparation method of the [ 18F]-labeled Larotrectinib compound includes

the following steps:

[0010] step 1: hydroxyl protection: protection of the active hydroxyl group of the

iodo-Larotrectinib analog precursor (I-Larotrectinib);

[0011] step 2: preparation of a labeled precursor: carrying out iodine activation on

Jodo-Larotrectinib analogue precursor (I-Larotrectinib) to prepare a trifluoroacetic acid

I-Larotrectinib analogue, and then causing the trifluoroacetic acid I-Larotrectinib analogue

to react with an adamantane-substituted auxiliary acid to prepare a labeled precursor: hydroxy-producted spirocyclic hypervalent I(III)-Larotrectinib analogue ; and

[0012] step 3: preparation of1 8 F-labeled product: carrying out substitution reaction

of the labeled precursor with a1 8 F radioactive source to prepare hydroxyl-protected

18F-Larotrectinib and synthesize a 18F-Larotrectinib analog, and then carrying out

deprotection to obtain 1 8 F-Larotrectinib and a1 8 F-Larotrectinib analogue.

[0013] The synthesis route of 18F-2-Larotrectinib is as follows:

F F

OH /N--N OOCC 6 H5 NN

N NiN 1 2 0 C 28H26FIN 60 3:640.46 C 2 1H 22FIN 60 2: 536.35 I - 2-Larotrectinib I - Bz-2-Larotrectinib

F F

F N- OOCCH5 0 /N. -N OOCCH 5 F3 CCOO H O F CCO'JN N6N!4 K)-, - N

C 3 2 H 2 FylN 6 O7 :868.50 H C4 1H 4 2 FIN0 7 :876.72 I - Bz-TfAc-2-Larotrectinib I(III)-SPIAd - Bz-2-Larotrectinib

F F

OOCC6H5 P N,-N OH F N 18 18 F N F N J~N rO CCH HN OH,

0 0 8 8 C 2 1H2 6F' FN60 3: 531.55 C 2 1H22F' FN 6 0 2 : 427.45 18 18 F -Bz-2-Larotrectinib F - 2-Larotrectinib

[0014] The synthesis route of 18F-5-Larotrectinib is the same as that of

18 F-2-Larotrectinib except that the substitution of I in the iodo-Larotrectinib analogue

precursor (I-Larotrectinib) in the raw material is 5-position substitution. The specific

synthesis route is as follows:

N OH N- N OOCCH, F F

FN N N N FN6

C 21H22FIN 602 : 536.35 C 28H 26FINS0 3:640.46 1-5- Larotrectinib I - Bz-5-Larotrectinib

H _ H

FCCOO O HO '-OOCCF 3 O H

NN OOCC 6Hs 0

-. N OOCC 6H

3 H N N6

C 32H 28FyN 6 07:868.50 4 1 - Bz-TfAc-5-Larotrectinib C 41H-42 FIN6 07:876.72 |(Ill)-SPIAd - Bz-5-Larotrectinib

18 18 F F

NN OH F N OOCC6H5

FN F N N N

8 18 C281H 26F FN60 3: 531.55 C21H 22F' FN 602 : 427.45 18 18 F -Bz-5-Larotrectinib F -5-Larotrectinib

[0015] The iodo-Larotrectinib analogue precursor in step 1 has the following

structural formula:

R2 N'N OR 3

-()N N N

[0016] in the above formula, RI and R2 each independently represent a phenyl

substituent, RI and R2 are each independently selected from group consisting of H,

halogen, hydroxyl, nitro, alkyl with 1 to 6 carbon atoms, alkenyl with 2 to 6 carbon atoms, alkynyl with 2 to 6 carbon atoms, cycloalkyl with 3 to 10 carbon atoms, aryl with 6 to 10 carbon atoms, heterocycloalkyl with 4 to 10 carbon atoms, and heteroaryl with 6 to 10 carbon atoms; RI and R2 each independently form a group with R3;

[0017] in the above formula, R3 specifically represents H; the protection of the

active hydroxyl functional group means that through esterification or etherification, R3 is

substituted with the following functional groups to protect the active hydroxyl, and the

substituted functional groups include trimethylsiloxane (TMS), tert-butyldimethylsiloxane

(TBDMS), triethylsiloxane (TES), tert-butyldiphenylsiloxane (TBDPS), methyl ether

(Me), benzyl ether (Bn), trityl ether (Tr), p-methoxytrityl ether (MMT), dimethoxytrityl

ether (DMT), tert-butyl ether ( tBu), methoxy methyl ether (MOM), 2-methoxy ethoxy

methyl ether (MEM), methylthio methyl ether (MTM), benzyloxy methyl ether (BOM),

p-methoxybenzyl ether (PMB), p-methoxybenzyloxy methyl ether (PMBOM),

3,4-dimethoxybenzyl ether (DMB), tetrahydropyran ether (THP), methoxycarbonyl (Moc),

ethoxycarbonyl (Eoc), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz),

9-fluorenylmethoxycarbonyl (Fmoc), acetyl (Ac), trifluoroacetyl (TfAc), chloroacetyl

Base (CAc), dichloroacetyl (DCAc), benzoyl (Bz), pivaloyl (Pv), methanesulfonyl (Ms),

benzylsulfonyl (Bs), allylsulfonyl (Bs), allyl Alkyloxycarbonyl (Als), allyloxycarbonyl,

alkanoyl with 1 to 16 carbon atoms,enoyl with 2 to 16 carbon atoms, alkynyl with 3 to 6

carbon atoms, cycloalkyl with 4 to 10 carbon atoms, aroyl with 7 to 16 carbon atoms, and

heterocycloalkanoyl with 4 to 10 carbon atoms; optionally, the substituted functional

groups can be substituted by 1, 2, 3 or 4 independent R3 groups, the alkanoyl is preferably

acetyl, and the aroyl is preferably benzoyl; the substituted functional groups can be

optionally substituted by 1, 2, 3 or 4 independent R3 groups, and the substituted alkanoyl

is pivaloyl or phenylacetyl.

[0018] The preparation of trifluoroacetic acid iodo-Larotrectinib analogue in step 2

is to cause the iodo-Larotrectinib analog precursor (I-Larotrectinib) with protected active hydroxyl prepared in step 1 to react with trifluoroacetic acid or trifluoroacetic anhydride, an organic solvent and an oxidant so that I is activated by trifluoroacetate;

[0019] the organic solvent includes one or more of chloroform, dichloromethane,

acetonitrile, acetone, and tert-butanol peroxide;

[0020] the oxidant includes a urea-hydrogen peroxide complex, Oxone,

2,2,6,6-tetramethylpiperidine-oxideormCPBA.

[0021] The adamantane-substituted auxiliary acid in step 2 is SPIAd, and the

conditions for the reaction of SPIAd with trifluoroacetic acid iodo-Larotrectinib analog are

as follows: SPIAd is mixed with one or more of sodium carbonate solution, MeCN,

NaHCO3 and acetone at 0°C, and the pH of the mixed solution is controlled to be 8 to 10,

and the mixed solution reacts with the trifluoroacetic acid iodo-Larotrectinib analogue at

°C while continuously stirring for 1 hour to 10 hours, thus obtaining the labeled

precursor.

[0022] The 18F radioactive source in step 3 is obtained by the following method:

[0023] (1) preparation of [ 18F] fluoride target water: producing [ 18F] fluoride target

water by 1 80(p,n) 18F nuclear reaction, and delivering the [ 18F] fluoride target water to a

sterile lead-protected hot cell of 18 0-enriched water by nitrogen pressure, wherein [ 18F]

fluoride target water produced by this method is usually further diluted with Milli-Q

(Millipore ultrapure water instrument) ultra-purified water to obtain 1-3mCi/ml [ 18F]

fluoride target water liquid before being used in research.

[0024] (2) enrichment of [ 18F] fluoride by QMA anion exchange solid phase

extraction cartridge (QMA): causing an aliquot of the target water containing an

appropriate amount of [ 18F] fluoride to slowly pass through the anion exchange solid

phase extraction cartridge (QMA) under a nitrogen flow, wherein the anion exchange solid

phase extraction cartridge (QMA) is pre-activated by washing with NaHCO 3(aq)(8.4%

1mL) and water (20 mL, until the pH indicator shows neutrality); enriching the 1[ 8 F]

fluoride on the QMA anion exchange solid phase extraction cartridge (QMA), and

removing 180 and other impurities by separation and elution, thus obtaining [1F] fluorine

source for the [ 18F] fluoride QMA anion exchange solid phase extraction cartridge

(QMA);

[0025] (3) eluting the [ 18F] fluoride enriched on the QMA anion exchange solid

phase extraction cartridge (QMA) to obtain a quaternary ammonium salt or inorganic salt

solution of [ 18F] fluoride; washing the [ 18F] fluoride enriched on QMA anion exchange

solid phase extraction cartridge (QMA) with a washing solution obtained by dissolving an

organic or inorganic base (for example, a certain amount of tetraethylammonium

bicarbonate, e.g., 8mg) in acetonitrile and water (lmL, v/v 7:3) or acetonitrile (ImL) or

methanol (lmL) or ethanol (lmL) and eluting the [ 18F] fluoride into a V-shaped flask

sealed with a Teflon-lined diaphragm, thus obtaining an acetonitrile aqueous solution or

an acetonitrile or methanol solution of the organic or inorganic salt of [ 18F] fluoride; and

[0026] (4) preparation of a dry quaternary ammonium salt or inorganic salt of [ 18F]

fluoride: heating the V-shaped flask containing the acetonitrile aqueous solution or

acetonitrile or methanol or ethanol solution of the organic or inorganic salt of [ 18F]

fluoride and sealed with a Teflon-lined diaphragm to 95 C to 110 'C, and at the same

time drying nitrogen gas by passing through a P20-DrieriteTM column to blow the

V-shaped flask, and then discharging the exhaust gas through the vented flask; when no

liquid is seen in the flask, taking the flask out of the hot bath, dosing anhydrous

acetonitrile (1 mL) in the flask, and heating the flask again until the flask is dry; repeating

this step another three times; then, cooling the flask at room temperature under nitrogen

flow, thus obtaining a dry organic or inorganic salt of [ 18F] fluoride, such as

[1 8F]KF/K2C03/K2.2.2, [1 8F]KF/K2C204/K2.2.2, [1 8F]KF/KOTf, [ 18F]Et4NF, [ 18F]Et4NHCO3,

[ 18F]Et4NOMs, and [ 18F]Et4NOTf; its radioactive [ 18F] fluoride recovery rate varies depending on the elution process used.

[0027] The substitution reaction between the labeled precursor and the 18F

radioactive source in Step 3: dosing a solvent in the V-shaped flask containing the dry

organic or inorganic salt of [ 18F] fluoride to re-dissolve the dry organic or inorganic salt,

and then dosing the labeled precursor to take reaction, thus obtaining a crude reaction

solution of undeprotected labeled product [1 8F]-Larotrectinib.

[0028] Deprotection in step 3 is implemented by the following method: dosing or

not dosing a certain amount of organic base or inorganic base or organic acid or inorganic

acid in the crude reaction solution of undeprotected labeled product [ 18F]-Larotrectinib,

and removing the hydroxyl protecting group by heating, thus obtaining a crude reaction

solution of labeled product [1 8F]-Larotrectinib.

[0029] The crude reaction solution of labeled product [ 18F]-Larotrectinib is

separated and purified by the following method: purifying the crude reaction solution of

labeled product [18F]-Larotrectinib by semi-preparative HPLC or Waters Sep-Pak C-18

cartridge, and rinsing the purified product with a solvent into a sterile vacuum flask, blow

drying with nitrogen at 60 °C for 20 minutes, and reconstituting a solution with brine,

wherein the obtain solution includes 100ul of 25% Vitamin C aqueous solution and 100ul

of 20% Tween 80 ethanol solution is a labeled product [ 18F]-2-Larotrectinib injection.The

labeled product [18F]-Larotrectinib injection is analyzed and identified by the following

method:

[0030] The identity and purity (radiochemical purity and chemical purity) of the

product are determined by radio-HPLC (60:40 CH3CN:H20+0.iN ammonium formate,

Phenomenex Luna Ci, 250 x 4.6 mm, 5 m, UV at 254 nm; CH3CN/0.1 M NH4•HCO2

(aq) (v/v, 7/3), flow rate: 1.0 mL/min) and radio-TLC (EtOAc + 0-5% EtOH). The product

had the radiochemical purity and chemical purity of greater than 90% to 99%. The

radiochemical yield is determined as the percentage of radioactivity of the labeled precursor dosed to the DMF-diluted [1 8 F]Et4NHCO 3 and separated as the final product from the amount of activity in the V-shaped reaction flask, without attenuation correction.

The radiochemical yield was 20 to 45.3 (without attenuation correction), the

radiochemical purity was greater than 99%, and the specific activity was 2.56 Ci/mol to

18 Ci/umol.

[0031] Compared with the prior art, the invention has the following beneficial

technical effects:

[0032] (1) The invention provides a1 8 F-labeled compound 1 8 F-Larotrectinib and

its analogs, and further provides a preparation method of the1 8 F-labeled compound and its

analogs; that is, the 1 8 F-labeled compound 18F-Larotrcetinib is prepared by the trivalent

iodine substitution method. The reaction speed is high, the reaction conditions are

relatively mild, the operation is simple, the reaction time is short, the post-processing is

simple, and the carrier-free radiolabeled compound can be prepared with a high

radiochemical purity.

[0033] (2) The invention provides a 18F-labeled compound 1 8F-Larotrectinib and

its analogues having the characteristics of emitting positrons; with the help of PET-CT

positron emission tomography technology, the distribution of the Larotrectinib compound

and its analogues in vivo and in tumors is visually displayed, and a new imaging agent is

provided for early tumor diagnosis.

DETAILED DESCRIPTION OF THE INVENTION

[0034] The following describes the embodiments of the present invention in detail.

The embodiments are implemented on the premise of the technical solutions of the present

invention. Detailed implementations and specific operation processes are given. However,

the protection scope of the present invention is not limited to the following embodiments.

[0035] In the following embodiments, various processes and methods that are not described in detail are conventional methods known in the art. Unless otherwise specified, materials, reagents, devices, instruments, equipment, etc. used in the following examples can be commercially available.

[0036] I-Larotrectinib analogue precursor in step 1

RR N-N OR 3

R) N N I0 oHN-,

[0037] In the above formula, R3 specifically represents H.

[0038] It can be prepared by the following method:

[0039] Step 1: key intermediate 4 and its synthesis

R2R 0 2 0I 02 00 R HN R ROH NH 2

0 " 0 -

. Ri R R1 R1 1 2 3 4

[0040] step 2: key intermediate 8 and its synthesis

- NN N

CI N C1 N NO 2 6 NH 2

[0041] Step 3: Preparation of halogenated larotrectinib analog 9

OH R1 N-N OH

CI N R1cN HN N N O0

[0042] Taking the preparation of (R)-2-F-5-I-Larotrectinib as an example, the

preparation method is as follows:

[0043] step 1: key intermediate 4 and its synthesis

[0044] 1a. Synthesis of (E)-4-(5-fluoro-2-iodophenyl)-4-oxobut-2-enoic acid

1 0 1 0 ' OH

10 0

F F C 8 H 6FIO: 264.04 C 1 0H 6 F103 : 320.06

[0045] 5-fluoro-2-iodoacetophenone (39.6g, 150mmol) and glyoxylic acid

monohydrate (13.9g, 151mmol) were dosed in a 500mL reactor, and then heated to react

and distilled off water under reduced pressure (95 °C, 0.1Mpa); after 3 hours of reaction,

the reaction mixture was cooled to room temperature, 5% potassium carbonate aqueous

solution (300mL) was then dosed, and the mixture was extracted twice with ethyl acetate,

200mL each time; after the aqueous layer was acidified (10% hydrochloric acid, 300mL),

and extraction was carried out twice with ethyl acetate, 200 mL each time; the organic

phases were combined, rinsed with brine, and dried over anhydrous sodium sulfate, and

the solvent was removed under reduced pressure, thus obtaining an orange solid. The solid

was dissolved in glacial acetic acid (50mL) and concentrated hydrochloric acid (36%,

mL), the mixture was heated to reflux for 4 hours, and the acetic acid was removed under

reduced pressure. The residue was extracted with ethyl acetate (300mL) and rinsed with

brine three times, 100mL each time; organic phase was dried over anhydrous sodium

sulfate, and the solvent was removed under reduced pressure, thus obtaining the target product (26.89g, 56%) as a yellow solid with an M.P. of 146 °C.

[0046] 'H NMR (400MHz, DMSO-d6) 6 12.96 (brs, 1H), 8.00-7.74 (m, 3H), 7.30

(t, 1H, J= 8.5 Hz), 6.64 (d, 1H, J= 15.4 Hz).

[0047] MS (El) m/z 320(M+)

[0048] lb. Synthesis of 4-(5-fluoro-2-iodophenyl)-4-oxobutanoic acid

1 0 1 0 - OH OH

0 - 0

F F C 1 0H 6FI03 :320.06 C1 0H 8 FI03 : 322.07

[0049] 210mL of acetic acid, 75mL of water, and starting material (47.1g,

147mmol) were dosed in a 500mL reaction flask; zinc powder (10.9g, 166mmol) was

dosed in the reaction mixture in batches within about 1 hour in a stirring way; the mixture

was further stirred for 3 hours, reactant was then filtered, the filter cake was rinsed with

ethyl acetate (300mL), the organic phase was rinsed with brine three times, 100mL each

time, and dried over anhydrous sodium sulfate, and the solvent was removed under

reduced pressure, thus obtaining the target product (29.8g, 63%) with an M.P. of 152 °C.

[0050] 'H NMR (400 MHz, CDCl3) 12.00 (1H, brs), 7.91-7.71 (3H, m), 3.23 (2H,

t, J= 6.26), 2.57 (2H, t, J = 6.24).

[0051] MS (El) m/z 322 (M+).

[0052] 1c. Synthesis of methyl 4-(5-fluoro-2-iodophenyl)-4,4-dimethoxybutanoate

1 0 I I 0 0 OH 0_

0 0

F F C1 0H 8 FI03: 322.07 C 13 H 1 6FI04 : 382.17

[0053] The starting material (32.2g, 0.1mol), trimethyl orthoformate (C4H1003=

106.12, 31.84g, 0.3mol) and methanol (90mL) were dosed in a 250mL reaction flask, and

drops of sulfuric acid was then dosed; the mixture was heated to 65 °C to react for 4 hours, TLC tracked the reaction process until the starting material was completely converted, and the solvent was distilled off under reduced pressure. The residue was diluted with isopropyl ether (190 mL), the reaction was quenched with saturated sodium bicarbonate (100 mL), the organic phase was separated, rinsed with brine twice, 120 mL each time, and dried over anhydrous magnesium sulfate, and the solvent was distilled off, thus obtaining the product (32.48g, 85%) which was directly used in the next step.

[0054] 'H NMR (400 MHz, CDC3) 1.29-1.38 (2H, m), 2.25 (2H, t, J = 7.2 Hz),

3.11 (3H, s), 3.17 (6H, s), 7.24-7.28 (1H, m), 7.31-7.38 (2H, m), 7.46 (1H, dt, J = 8.6, 1.4

Hz);

[0055] MS (ESI) m/z 383 [M+H]*.

[0056] 1d. Synthesis of 4-(5-fluoro-2-iodophenyl)-4-oxobutanamide

II I I I I 00 100 O- NH 2

0 0

F F C 13H 1 6FI04 : 382.17 C 12 H 15FIN03 : 367.16

[0057] 100mL of methanol was dosed in a 250mL reaction flask and cooled to

below 0 °C, and ammonia gas was introduced until the solution was saturated (about

-12g); the esterification product (19.lg, 0.05mol) from step I was dosed in 50mL of

methanol to obtain a liquid, the liquid was dropwise dosed in the methanol-ammonia

saturated solution, and the internal temperature was maintained at about 0 °C. After the

liquid was completely dosed dropwise, the reaction solution continued to react for 16

hours at 0 °C. When TLC detected that the esterification product from step I in the

reaction solution basically disappeared, the reaction was stopped, and the solvent was

removed under reduced pressure, thus obtaining an oily product which was used directly

in the next step of reaction.

[0058] MS (ESI) m/z: 368.1(M+H)+.

[0059] le. Synthesis of 5-(5-fluoro-2-iodophenyl)-3,4-dihydro-2H-pyrrole

1 0 I I I N (E) NH12

0 F F C 12H 15 FIN03 :367.16 C 10 H 9FIN: 289.09

[0060] Dry tetrahydrofuran (300mL), the amide (18.36g, 0.05mol) from step Id,

and sodium borohydride (29.26g, 0.77mol) were dosed in a 500mL reactor, stirred to be

uniform, and then cooled to 0 °C with an ice bath; in the presence of nitrogen, boron

trifluoride ether solution (36.75mL, 0.3mol) was dosed dropwise within about 2 hours, the

ice bath was removed, the mixture was heated to reflux for 16 hours, TLC detected the

reaction until raw material was completely converted, the reaction solution was cooled to

°C, 6N hydrochloric acid (35mL) was slowly dosed dropwise, the mixture was heated to

reflux for 1 hour, the reaction solution was cooled to 40 °C, and the solvent was removed

under reduced pressure. The residue was diluted with water, neutralized with 10% NaOH

to neutrality, extracted with chloroform, dried over anhydrous sodium sulfate, filtered and

concentrated under reduced pressure, thus obtaining oily

-(5-fluoro-2-iodophenyl)-3,4-dihydro-2H-pyrrole(11.27g,78%).

[0061] The NMR data of the product obtained were as follows:

[0062] 'H NMR, 400 MHz, CD30D 6: 7.78 (m, 1H), 7.43-7.34 (m, 2H), 3.54 (m,

2H), 2.12 (dt, J = 10.3, 2.0 Hz, 2H), 1.97 (dt, J = 15.8, 7.9 Hz, 2H).

[0063] 13C NMR, 100MHz, CD30D 6: 176.2, 158.2 (d, J = 260.3), 155.0 (d, J=

10.3), 124.0 (d, J= 3.1), 119.4 (d, J= 23.5), 118.4 (d, J= 23.5), 62.3, 36.5, 21.0.

[0064] If: Synthesis of 2-(5-fluoro-2-iodophenyl)pyrrolidine

I N I HN (E) j

F F C 10 H9 FIN: 289.09 C 10 H 11 FIN: 291.11

[0065] The oily 5-(5-fluoro-2-iodophenyl)-3,4-dihydro-2H-pyrrole (7.23g,

0.025mol) was dosed in 100mL of a solution of methanol and water (4:1); the mixed

solution was cooled to 0 °C, sodium borohydride (0.95g, 0.025mol) was dosed in batches,

hydrogen was released, and the reaction solution turned into a yellow turbid liquid; 3

hours later, the reaction solution was heated to room temperature, and the solvent was

distilled off. The residue was treated with NaOH and then extracted with isopropyl ether;

the ether layer was dried over anhydrous sodium sulfate overnight and filtered to remove

the solvent, thus obtaining a light yellow oily product (6.23g, 88.3%), which was directly

used into next step of reaction.

[0066] 'H NMR, 400 MHz, CDCl3 6: 7.29 (m, 1H), 7.02 (m, 1H), 6.96 (m, 1H),

4.09 (t, J = 7.8 Hz, 1H), 3.16(m, 1H), 3.04(m, 1H), 2.21-2.30(m, 1H), 1.77-1.95(m, 3H),

1.57-1.67(m, 1H).

[0067] LC-ESI-MS (m/z) 292 [M+H]*.

[0068] Step 2: Synthesis of key intermediate 8

[0069] 2a. Synthesis of 5-chloropyrazolo[1,5-a]pyrimidin-3-amine

5:0'N- N.N - N

CI N CI N NO 2 NH 2

C6H 3 CIN 40 2 : 198.57 C 6 H 5 CIN 4 : 168.58

[0070] 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine compound (25g, 0.125mol),

ethanol (250mL), and iron powder (75g, 1.25mol) were dosed in a 1000mL reaction flask

and heated to reflux in the presence of nitrogen, and ammonium chloride (66.5g, 1.25mol)

aqueous solution (250mL) was dosed dropwise at the same time within about 1 hour; the

reflux reaction was continued for 6 hours; TLC tracked the reaction until the reaction was

completed, the reactant was concentrated to paste under reduced pressure. The residue was

diluted with water (100mL), then layered with dichloromethane and extracted 4-6 times,

200mL each time; the organic phases were combined, rinsed with saturated brine, dried

over anhydrous magnesium sulfate, and concentrated under reduced pressure, thus

obtaining 5-chloropyrazolo[1,5-a]pyrimidin-3-amine compound (419.7g, 93.5%).

[0071] 'H NMR, 400 MHz, CD30D 6:9.29(d,J= 7.2Hz,1H),8.71(s,1H),8.16(d,J

7.2Hz,1H), 5.92(s,2H):

[0072] MS(ESI)m/z: 169.8[M+H]+.

[0073] 2b. Synthesis of 5-chloro-3-isocyanatopyrazolo[1,5-a]pyrimidine

C1 N C1 N NH 2 N-CZO C 6H 5CIN 4 : 168.58 C 7H 3CIN 4 0:194.58

[0074] Triphosgene (9.91g, 33.38mmol) was dissolved in 50ml of tetrahydrofuran,

-chloropyrazolo [1,5-a]pyrimidin-3-amine (16.86g, 0.1mol) and triethylamine (0.47g,

4.64mmol) were dosed, and the reaction system was stirred to react for 1 hour at 25°C. A

suspension of 5-chloro-3-isocyanatopyrazolo[1,5-a]pyrimidine was obtained, and the

product was directly subjected to the next reaction without purification.

[0075] 2c. Synthesis of

(S)-N-(5-chloropyrazolo[1,5-a]pyrimidin-3-yl)-3-hydroxypyrrolidine-1-carboxamide

- N OH

N~ - ~ NI

C1CI N N6 CI N- HN NC::O 6 O

C7 H 3 CIN 4 0:194.58 C 11 H 12 CIN 5 02 :281.70

[0076] Sodium bicarbonate solution (80ml, 0.5M, 40mmol) was dosed in a 250mL

reactor, (S)-pyrrol-3-ol (79g, 55mmol) was then dosed, the isocyanate solution prepared in

step 9b (62mL, 100mmol, calculated on the basis of 5 -chloropyrazolo

[1,5-a]pyrimidin-3-amine) was dosed dropwise, and the temperature was maintained at

about 0 °C; and then the mixture was further stirred at 0 °C for 3 hours and then heated to

°C to further react for 5 hours; the reaction solution was cooled to room temperature,

extracted three times with ethyl acetate, 100 mL each time. The organic phases were

combined, rinsed with 2N hydrochloric acid, then with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography and elution with ethyl acetate/petroleum ether (5:5, v/v) was carried out, thus obtaining a product (22g, 78%).

[0077] 'H NMR(300MHz,d6DMSO)6 : 9.26(d,J = 7.2Hz 1H), 8.73(s,1H),

8.78(s,1H), 8.07(d,J= 7.2Hz,1H),4.02(m,1H), 3.62(m,2H),3.48(m,2H),2.36(m,2H);

[0078] MS(ESI)m/z: 282[M+H]+.

[0079] Step 3: Preparation of halogenated larotrectinib analog 9

[0080] Synthesis of halogenated larotrectinib compound (R)-5-F-2-I-Larotrectinib

F - NN OH I HN / N OH

C NI N + | NH HN-I(NHN N

C 11H 12CIN 502 :281.70 C 10H 11FIN:291.11 C 21H22 FIN 602: 536.35 2-(5-fluoro-2-iodophenyl)pyrrolidine (R)-5-F-2-1-Larotrectinib

[0081] Pyrazolo (1,5a) pyrimidine compound (30g, 106mmol), 5-F-2-I-pyrrole

compound (30.9g, 106mmol) and 1,4-dioxane (150mL) were dosed in a reaction flask,

N,N-lutidine (C7H1ON2=122.17, 25.9g, 212mmol) was dosed dropwise, and the reaction

temperature was maintained within 30 °C; then, the reaction was continued for 6 hours and

the whole reaction was under the control of TLC; the post treatment was carried out in the

same way as that in Example 6, and the racemic mixture 5-F-2-I-Larotrectinib (53.8g,

94.6%) was obtained as a white solid. Then, by chiral LC separation and purification,

(R)-5-F-2-I-Larotrectinib was obtained as a pure white chiral solid, and

(S)-5-F-2-I-Larotrectinib was obtained as a pure white chiral solid.

[0082] 1H NMR(300MHz,d 6DMSO) 6 = 9.12(d,J = 7.2Hz 1H), 8.73(s,1H),

8.78(s,1H),

8.07(d,J=7.2Hz,1H),6.8-7.3(m,3H),4.17(m,1H),4.02(m,1H),3.62(m,2H),3.48(m,2H),1.75

2.86(m,8H);

[0083] MS(ESI)m/z: 536.3(M+H)+.

[0084] The preparation process of the compound of the invention will be described

in detail below by examples, where

[0085] Examples 1 to 42 describe hydroxyl protection in the first step to prepare I

hydroxyl protected Larotrectinib compound

[0086] Example 1

[0087] Preparation of I-Bz-2-Larotrectinib or I-Bz-5-Larotrectinib by benzoyl

chloride esterification

[0088] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

were dosed in a 2000 ml reaction flask and stirred at room temperature to be dissolved;

pyridine (240mmol, 18.98g) and benzoyl chloride (80mmol, 11.4g) were dosed in the

solution at the same time; the mixture was stirred at room temperature to react for 3 hours

under the control of TLC; at the end of the reaction, the mixture was poured into water and

the organic layer was separated; the organic layer was sequentially washed with dilute HCl

to remove pyridine and acidify unreacted acid chloride), with dilute NaOH (preferably

saturated sodium bicarbonate, lOOmL) to wash off acid, and then with saturated sodium

chloride to neutral, and most of the water in the organic layer was removed. The organic

layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to

remove the solvent, and the residue was crystallized with ethanol-methyl tert-butyl ether

(4:1, 180mL), thus obtaining the product I-Bz-2-Larotrectinib as a white solid with a yield

of 88% (70.4mmol, 45.lg). The synthesis route was as follows: F F

N H 00 QN NJ~

C 21H 22FIN 60 2: 536.35 C28H 26FIN0 3:640.46 1 -2- Larotrectinib I - Bz-2-Larotrectinib

[0089] Example 2

[0090] Preparation of I-Bz-5-Larotrectinib by benzoyl chloride esterification

[0091] The preparation method was the same as that in Example 1. The compound

1-5-Larotrectinib was used as the raw material to obtain the product I-Bz-5-Larotrectinib

as a white solid with a yield of 88% (70.4mmol, 45.1g). The route was as follows:

I0

F F N HNF N N F NN N N

C 21H 22FIN 602: 536.35 C28H 26FIN 603:640.46 I -5- Larotrectinib I - Bz-5-Larotrectinib

[0092] Example 3

[0093] Preparation of I-Piv-2-Larotrectinib by pivaloyl chloride esterification

[0094] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

saturated sodium bicarbonate solution (23g) and pivaloyl chloride (80mmol, 9.64g) were

dosed in the solution at the same time; the mixture was stirred at room temperature to

react for 24 hours under the control of TLC; at the end of the reaction, the mixture was

poured into water and the organic layer was separated; the organic layer was sequentially

washed with IN sodium hydroxide (1OOmL) and saturated brine (1OOmL), dried over

anhydrous magnesium sulfate, and concentrated under reduced pressure to remove the

solvent; the residue was crystallized with methanol-isopropyl ether (7:3, 150 mL), thus

obtaining the product I-Piv-2-Larotrectinib as a white solid with a yield of 85% (68mmol,

42.2g). The route was as follows:

F F PN-N OH N'-N

N NN N6

C 21H 22FIN 60 2:536.35 C 26H30FIN 60 3:620.47 I - 2-Larotrectinib I - Piv-2-Larotrectinib

[0095] Example 4

[0096] Preparation of I-Piv-5-Larotrectinib by pivaloyl chloride esterification

[0097] The preparation method was the same as that in Example 3. The compound

1-5-Larotrectinib was used as the raw material to obtain the product I-Piv-5-Larotrectinib

as a white solid with a yield of 85% (70.4mmol, 42.2g). The route was as follows:

II 0

N'.N 0 F P'N-N OH OHF

FNN N

C 21H22FIN 60 2: 536.35 C 26H 30FING0 3:620.47 I- 5-Larotrectinib I - Piv-5-Larotrectinib

[0098] Example 5

[0099] Preparation of I-Ac-2-Larotrectinib by acetyl chloride esterification

[00100] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

triethylamine (240mmol, 24.2g) and acetyl chloride (80mmol, 6.28g) were dosed in the

the organic layer was separated; the organic layer was sequentially washed with saturated

ammonium chloride solution (100mL), saturated sodium bicarbonate, and water, dried

over anhydrous magnesium sulfate, and concentrated under reduced pressure to remove

the solvent; the residue was crystallized with acetone-petroleum ether (5:2, 110 mL), thus obtaining the product I-Ac-2-Larotrectinib as a white solid with a yield of 96.9%

(77.52mmol, 44.83g).

F F OH N-N NN

N N6 N

C 21H 22 FIN 6 0 2 : 536.35 C23 H24FING0 3:578.39 I - 2-Larotrectinib I - Ac-2-Larotrectinib

[00101] Example 6

[00102] Preparation of I-Ac-5-Larotrectinib by acetyl chloride esterification

[00103] The preparation method was the same as that in Example 5. The compound

1-5-Larotrectinib was used as the raw material to obtain the product I-Ac-5-Larotrectinib

as a white solid with a yield of 96.5% (77.2mmol, 44.7g). The route was as follows:

OH OH N.. 0 roN - N F N N F

16 o

C 21H22FIN 602 : 536.35 C23H24FING0 3:578.39 I - 5-Larotrectinib I - Ac-5-Larotrectinib

[00104] Example 7

[00105] Preparation of I-Tf-2-Larotrectinib by trifluoroacetic anhydride

esterification

[00106] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

2,4,6-trimethylpyridine (240mmol, 29.1g) as a base and trifluoroacetic anhydride

(80mmol, 16.8g) were dosed in the solution at the same time; the mixture was stirred at

room temperature to react for 8 hours under the control of TLC; at the end of the reaction,

the mixture was poured into water and the organic layer was separated; the organic layer

was sequentially washed with 5% dilute hydrochloric acid solution (1OOmL) and saturated

brine (100mL), dried over anhydrous magnesium sulfate, and concentrated under reduced

pressure to remove the solvent; the residue was crystallized with isopropyl

acetate-petroleum ether (6:1, 120mL), thus obtaining the product I-Tf-2-Larotrectinib as a

white solid with a yield of 94% (75.2mmol, 59.44g). The route was as follows:

F F

NN O CF N-N OH I CF3

NN N N6

1 2 0 C 21H22FIN 60 2: 536.35 C 23H 21F 41N 60 3:632.36 I - 2-Larotrectinib I -Tf-2-Larotrectinib

[00107] Example 8

[00108] Preparation of I-Tf-5-Larotrectinib by trifluoroacetic anhydride

esterification

[00109] The preparation method was the same as that in Example 7. The compound

1-5-Larotrectinib was used as the raw material to obtain the product I-Tf-5-Larotrectinib as

a white solid with a yield of 94% (75.2mmol, 59.4g). The route was as follows:

NN OH N.N O CF F FCF F NN FN N

1 02 0 C21 H22 FIN 6 02 : 536.35 C 23H 21F 4 1NG0 3:632.36 I -5-Larotrectinib I - Tf-5-Larotrectinib

[00110] Example 9

[00111] Preparation of I-DCAc-2-Larotrectinib by dichloroacetyl chloride

esterification

[00112] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

pyridine (240mmol, 19.0g) and dichloroacetyl chloride (80mmol, 11.79g) were dosed in

the solution at the same time; the mixture was stirred at room temperature to react for 2

hours under the control of TLC; at the end of the reaction, the mixture was poured into

water and the organic layer was separated; the organic layer was sequentially washed with

saturated ammonium chloride solution (100mL), saturated sodium bicarbonate, and water,

dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to

remove the solvent; the residue was crystallized with acetone-petroleum ether (5:2, 110

mL), thus obtaining the product I-DCAc-2-Larotrectinib as a white solid with a yield of

91.6% (73.28mmol 47.43g). The route was as follows:

F F

N.-N OH N-N OOCCHC 2

N N HN N f N

0 0 C2 1H 2 2FIN60 2 :536.35 C23H 22Cl2FIN 6 0 3 :647.27 -2- Larotrectinib 1 I-DCAc -2- Larotrectinib

[00113] Example 10

[00114] Preparation of I-DCAc-5-Larotrectinib by dichloroacetyl chloride

esterification

[00115] The preparation method was the same as that in Example 9. The compound

1-5-Larotrectinib was used as the raw material to obtain the product

I-DCAc-5-Larotrectinib as a white solid with a yield of 91% (73mmol, 47g). The route

was as follows:

SN'N OH N' OOCCHCl 2

N N F N N

0 0 C 21H 22 FIN 6 02 : 536.35 C23H 22 CI2 FIN6 O 3 :647.27 1 -5- Larotrectinib I-DCAc -5- Larotrectinib

[00116] Example 11

[00117] Preparation of I-Moc-2-Larotrectinib by methyl chloroformate

esterification

[00118] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

pyridine (240mmol, 19.0g) and methyl chloroformate (80mmol, 47.2g) were dosed in the

solution at the same time; the mixture was stirred at room temperature to react for 8 hours

the organic layer was separated; the organic layer was washed twice with water,1OOmL

each time, dried over anhydrous magnesium sulfate, and concentrated under reduced

pressure to remove the solvent; the residue was crystallized with ethyl acetate-petroleum

ether (6:1, 140mL), thus obtaining the product I-Moc-2-Larotrectinib as a white solid with

a yield of 96.5% (77.2mmol, 45.88g). F F

N OH /N-N 0 O

NNN KN N6

1 -2

C 21H 22FIN 6 02: 536.35 25 C 23H 24FING0 3:594.39 I--2-Larotrectinib I - Moc-2-Larotrectinib

[00119] Example 12

[00120] Preparation of I-Moc-5-Larotrectinib by methyl chloroformate

esterification

[00121] The preparation method was the same as that in Example 11. The

compound 1-5-Larotrectinib was used as the raw material to obtain the product

I-Moc-5-Larotrectinib as a white solid with a yield of 91% (73mmol, 47g). The route was

as follows:

NN OH N-N OAO

F N N F N N( 1 2 0

C 21H22FIN 60 2: 536.35 C23 H24FINS0 3:594.39 I --5-Larotrectinib I - Moc-5-Larotrectinib

[00122] Example 13

[00123] Preparation of I-Eoc-2-Larotrectinib by ethyl chloroformate esterification

[00124] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

N,N-Diisopropylethylamine (240mmol, 30.48g) and ethyl chloroformate (80mmol, 8.68g)

were dosed in the solution at the same time; the mixture was stirred at room temperature

to react for 6 hours under the control of TLC; at the end of the reaction, the mixture was

washed with 5% dilute hydrochloric acid solution (100mL) and saturated brine (100mL),

remove the solvent; the residue was crystallized with isopropyl acetate-petroleum ether

(6:1, 120mL), thus obtaining the product I-Eoc-2-Larotrectinib as a white solid with a

yield of 94.3% (75.2mmol, 59.44g). The route was as follows:

F F 0

N OH NO O

N N N Nf

1 2 0

C 21H22FIN 602: 536.35 C 24H 26FIN 604 :608.41 I - 2-Larotrectinib I -2- Eoc-Larotrectinib

[00125] Example 14

[00126] Preparation of I-Eoc-5-Larotrectinib by ethyl chloroformate esterification

[00127] The preparation method was the same as that in Example 13. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-Eoc-5-Larotrectinib as a white solid with a yield of 94% (75mmol - 75g). The route was

as follows:

FF P'N-XF OH N O 0-\

1 02 0

C 21H22FIN 60 2: 536.35 C24 H2 6FIN 60 4:608.41 I - 5-Larotrectinib I - Eoc-5-Larotrectinib

[00128] Example 15

[00129] Preparation of I-Boc-2-Larotrectinib by Boc anhydride esterification

[00130] 400 mL of chloroform and compound1-2-Larotrectinib (80mmol, 42.9g)

DMAP (4mmol, 0.5g), KOH (240mmol, 13.44g), and Boc anhydride (80mmol, 17.46g)

to react for 16 hours under the control of TLC; at the end of the reaction, the mixture was

washed with 5% dilute hydrochloric acid solution (100mL) and saturated brine (100mL), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to remove the solvent; the residue was crystallized with isopropyl acetate-petroleum ether

(6:1, 120mL), thus obtaining the product I-Boc-2-Larotrectinib as a white solid with a

yield of 94% (75.2mmol, 47.86g). The route was as follows:

F F

NN OH N.-N OBoc

NN N6 NNN

C 21H 22FIN 602 : 536.35 C 26H 30FINS 4: 636.47 I - 2-Larotrectinib I - Boc-2-Larotrectinib

[00131] Example 16

[00132] Preparation of I-Boc-5-Larotrectinib by Boc anhydride esterification

[00133] The preparation method was the same as that in Example 15. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-Boc-5-Larotrectinib as a white solid with a yield of 94% (75mmol, 47g).

[00134] Example 17

[00135] Preparation of I-Troc-2-Larotrectinib by 2,2,2-trichloroethyl chloroformate

(Troc) esterification as the following route:

F F 0

N OH N-N CClI

N C lN NN

C 21H 22 FIN 6 0 2 : 536.35 C 24H 23Cl 3FIN 604:711.74 I - 2-Larotrectinib I -Troc-2-Larotrectinib

[00136] 150 mL of DCM, compound 1-2-Larotrectinib (80mmol, 42.91g) and

pyridine=79 (2.5eq, 200mmol, 15.80g) were dosed in a 500 ml reaction flask and stirred at

room temperature for 5 minutes; 2,2,2-trichloroethyl chloroformate (C3H2C1402=211.85,

1.2e, 96mmol, 13.66g, 20.34mmol) was dropwise dosed and dissolved in the DCM

(30ml) solution, and the solution was stirred at room temperature for 3 hours under the

control of TLC; at the end of the reaction, the mixture was poured into water and the

organic layer was separated; the organic layer was sequentially washed with 3% HCl and

saturated sodium bicarbonate and then washed three times with brine (80gx3); the organic

phase was dried over anhydrous sodium sulfate and filtered with suction, the filtrate was

concentrated to be viscous, and the residue was subjected to silica gel column

chromatography (petroleum ether: ethyl acetate = 5:1), thus obtaining the product

I-Troc-2-Larotrectinib as a white solid (91.2%, 51.93g).

[00137] Example 18

[00138] Preparation of I-Troc-5-Larotrectinib by 2,2,2-trichloroethyl chloroformate

(Troc) esterification

[00139] The preparation method was the same as that in Example 17. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-Boc-5-Larotrectinib as a white solid with a yield of 91% (72.8mmol 51g).

[00140] Example 19

[00141] Preparation of I-Teoc-2-Larotrectinib by trimethylsilyl ethoxycarbonyl

chloride (Teoc) esterification

F F 0

N OH .N 0 O N N N

N N3 0 0 C21 H 22 FIN 6 0 2 :536.35 C 27 H 34 FIN 60 4 Si:680.59 I -2-Larotrectinib I -Teoc-2-Larotrectinib

[00142] 150 mL of dichloromethane, compound1-2-Larotrectinib (80mmol, 42.91g)

and pyridine=79 (2.5eq, 200mmol, 15.80g) were dosed in a 500 ml reaction flask and cooled to 0-5 °C; trimethylsilyl ethoxycarbonyl chloride (Teoc) (C6H3ClO2Si= 180.70,

2.Oeq,28.91g, 160mmol) was dropwise dosed slowly and dissolved in the DCM (30ml)

solution; then, the obtained solution was further stirred at room temperature for 3 hours

under the control of TLC; at the end of the reaction, 5 mL of methanol was dosed and the

mixture was stirred to react for half an hour at room temperature; the mixture was poured

into water and the organic layer was separated; the organic layer was sequentially washed

with 3% HCl and saturated sodium bicarbonate and then washed three times with brine

(80gx3); the organic phase was dried over anhydrous sodium sulfate and filtered with

suction, the filtrate was concentrated to be viscous, and the residue was subjected to silica

gel column chromatography (petroleum ether: ethyl acetate = 5:1), thus obtaining the

product I-Teoc-2-Larotrectinib as a white solid (83.6%, 45.52g).

[00143] Example 20

[00144] Preparation of I-Teoc-5-Larotrectinib by trimethylsilyl ethoxycarbonyl

chloride (Teoc) esterification

[00145] The preparation method was the same as that in Example 19. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-Teoc-5-Larotrectinib as a white solid with a yield of 83% (66.4mmol, 45g).

[00146] Example 21

[00147] Preparation of I-TBS-2-Larotrectinib by tert-butyldimethylchlorosilane

etherification

F F "CN'N OTBS N-N OH

N N N N6

C27H3 FIN 60 2Si:650.61 C21 H 22 FIN 6 02 :536.35 I - 2-Larotrectinib I -TBS-2-Larotrectinib

[00148] 150 mL of DMF was dosed in a 500 ml reaction flask and cooled to 0 °C,

and then compound 1-2-Larotrectinib (80mmol, 42.91g), tert-butyldimethylchlorosilane

(TBS-Cl, C6H15ClSi= 150.72) (1.3eq, 104mml, 15.67g), and imidazole (C 3 H 4N 2 = 68.08,

1.5eq, 120mmol, 8.17g) were sequentially dosed; the mixture was stirred at room

temperature and reacted overnight usually until the conversion was complete under the

control of TLC; the mixture was diluted with an equal volume of chloroform, washed

three times with water and then washed once with brine. The organic layer was dried,

concentrated under reduced pressure, and the residue was subjected to silica gel column

chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the product

I-TBS-2-Larotrectinib (86.8%, 45.17g).

[00149] Example 22

[00150] Preparation of I-TBS-5-Larotrectinib by tert-butyldimethylchlorosilane

etherification

[00151] The preparation method was the same as that in Example 21. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-TBS-5-Larotrectinib as a white solid with a yield of 86% (68.8mmol, 45g).

[00152] Example 23

[00153] Preparation of I-TIPS-2-Larotrectinib by triisopropylchlorosilyl

etherification

F F N.N OTIPS .. N OH N N N N

C 21H22FIN 602 : 536.35 C30 H42 FIN 602Si:692.69 I - 2-Larotrectinib I - TIPS-2-Larotrectinib

[00154] 150 mL of DMF was dosed in a 500 ml reaction flask and cooled to 0 °C,

and then compound 1-2-Larotrectinib (80mmol, 42.91g), triisopropylchlorosilane (TIPSCl,

C9H21ClSi = 192.8, 1.3eq, 104mmol, 20.05g), and imidazole (C3H4N2 = 68.08, 1.3eq,

104mmol, 7.07g) were sequentially dosed; the mixture was stirred at room temperature

and reacted overnight usually until the conversion was complete under the control of TLC;

the mixture was diluted with an equal volume of chloroform, washed three times with

water and then washed once with brine. The organic layer was dried, concentrated under

reduced pressure, and the residue was subjected to silica gel column chromatography

(petroleum ether: ethyl acetate=5:1), thus obtaining the product I-TIPS-2-Larotrectinib

(88.8%, 49.21g).

[00155] Example 24

[00156] Preparation of I-TIPS-5-Larotrectinib by triisopropylchlorosilyl

etherification

[00157] The preparation method was the same as that in Example 23. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-TIPS-5-Larotrectinib as a white solid with a yield of 88% (70.4 mmol, 49g).

[00158] Example 25

[00159] Preparation of I-TBDPS-2-Larotrectinib by tert-Butyldiphenylchlorosilane

etherification F F

N OH N~N OTBDPS

N NTN N(

0 0

C 21H 22FIN 602 : 536.35 C 37H 40FIN 60 2Si:774.75 I - 2-Larotrectinib I - TBDPS-2-Larotrectinib

[00160] 100 mL of THF, compound 1-2-Larotrectinib (80mmol, 42.91g), and

imidazole (2eq, 160mmol, 10.89g) were sequentially dosed in a 500 ml reaction flask and

stirred to react for half an hour, and tert-butyldiphenylchlorosilane (TBDPSCl, 1.5eq,

120mmol, 32.98g) was then dosed in the solution; the mixture was stirred at 25 °C and

completely reacted under the control of TLC; the mixture was diluted with 300mL of

chloroform, and washed once with ammonium chloride solution (1OOmL, containing 37.2g of ammonium chloride), washed with saturated sodium bicarbonate twice, 100ml each time, and further washed once with brine (100ml), the organic phase was dried over anhydrous magnesium sulfate, and filtered; the mixture was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the product I-TBDPS-2-Larotrectinib (85.7%,

53.12g).

[00161] Example 26

[00162] Preparation of I-TBDPS-5-Larotrectinib by tert-Butyldiphenylchlorosilane

etherification

[00163] The preparation method was the same as that in Example 25. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-TBDPS-5-Larotrectinib as a white solid with a yield of 86% (68.8mmol 53g).

[00164] Example 27

[00165] Preparation of I-PMB-2-Larotrectinib by PMB-(p-Methoxybenzyl)ether

F F OMe

-XN XH .N O~

N- N NO P N NON

C 21H 22FIN 602 : 536.35 C29 H3 0FINS0 3:656.50 I - 2-Larotrectinib I - PMB-2-Larotrectinib

[00166] 100 mL of DMF and the dissolved solution of compound -2-Larotrectinib

(80mmol, 42.91g) were dosed in a 500 ml reaction flask; in the presence of nitrogen,

sodium hydrogen (1.3e, 104mmol, 2.5g) cooled to 0 °C was dissolved in the DMF (20mL)

solution; the mixture was stirred at 0 °C for 30 minutes; p-methoxybenzyl bromide (1.3eq,

104mmol, 201.0g) was dosed dropwise in the above solution, and then a catalytic amount

of crown ether was dosed; the reaction solution was further stirred at room temperature for

16 hours under the control of TLC till the end of the reaction; methanol was slowly dosed to quench the reaction; the solution was diluted with chloroform and water and then washed three times with water and twice with brine; the organic layer was dried over anhydrous sodium sulfate, and the mixture was concentrated under reduced pressure; the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the product I-PMB-2-Larotrectinib (82.8%, 44.15g).

[00167] Example 28

[00168] Preparation of I-PMB-5-Larotrectinib by P-methoxybenzyl etherification

(PMB-(p-Methoxybenzyl)ether)

[00169] The preparation method was the same as that in Example 27. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-PMB-5-Larotrectinib as a white solid with a yield of 83% (66.4mmol, 43.6g).

[00170] Example 29

[00171] Preparation of I-MOM-2-Larotrectinib by chloromethyl methyl ether

etherification

F F .- N OH / N-N OMOM

N NN N6

1i 2 0

C 21H 22FIN 6 02: 536.35 C 23H 26FIN 03 :580.40 I - 2-Larotrectinib I - MOM-2-Larotrectinib

[00172] 180mL of anhydrous DCM, compound1-2-Larotrectinib (80mmol, 42.91g),

and anhydrous N,N-diisopropylethylamine (2.0 eq, 160mmol, 20.68g) were dosed in a 500

mL reaction flask in the presence of nitrogen, and the reaction mixture was cooled to 0 °C;

chloromethyl methyl ether (MOM-Cl, 1.5eq, 120mmol, 9.66g) was dosed; the mixture

was heated to room temperature and stirred to react completely for about 8 hours or more

under the control of TLC; the reaction solution was washed once with ammonium chloride solution (100mL, containing 37.2g of ammonium chloride), washed twice with saturated sodium bicarbonate, 100ml each time, and washed once with brine (100ml); the organic phase was dried over anhydrous magnesium sulfate and filtered; the mixture was concentrated under reduced pressure; and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the product

I-MOM-2-Larotrectinib (74.4%, 34.54g).

[00173] Example 30

[00174] Preparation of I-MOM-5-Larotrectinib by chloromethyl methyl ether

etherification

[00175] The preparation method was the same as that in Example 29. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-MOM-5-Larotrectinib as a white solid with a yield of 74% (59mmol, 34g).

[00176] Example 31

[00177] Preparation of I-MEM-2-Larotrectinib by methoxyethoxymethyl chloride

etherification

F F N OH N'N OMEM N N N N

12

C 21H22FIN 602: 536.35 C2 5H30FIN0 4:624.46 1 -2- Larotrectinib I - MEM-2-Larotrectinib

[00178] 180mL of dry DCM, compound 1-2-Larotrectinib (0.8mmol), and

methoxyethoxymethyl chloride (MEM-Cl, 1.5eq, 120mmol, 14.95g) were dosed in a 500

mL reaction flask, and N, N-Diisopropylethylamine (1.5eq, 120mmol, 15.51g) was then

dosed; the reaction mixture was stirred at room temperature and completely reacted for

about 5 hours under the control of TLC; the reaction mixture was diluted with DCM

(180mL), and the obtained mixture was then washed twice with water, 100ml each time and then washed with brine (100ml); the organic phase was dried over anhydrous magnesium sulfate, and filtered; the mixture was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the product I-MEM-2-Larotrectinib (76.8%, 38.37g).

[00179] Example 32

[00180] Preparation of I-MEM-2-Larotrectinib by methoxyethoxymethyl chloride

etherification

[00181] The preparation method was the same as that in Example 30. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-MEM-5-Larotrectinib as a white solid with a yield of 76% (60.8mmol - 38g).

[00182] Example 33

[00183] Preparation of I-SEM-2-Larotrectinib by 2-(trimethylsilyl)ethoxymethyl

chlorideetherification

F F

Z;"N-N OH N..-N 0 O

N N OH0 N 0 N6 0

C21 H22 FIN 6 02 : 536.35 C27 H 36FIN 6 0 3 Si:666.61 I - 2-Larotrectinib I -SEM-2-Larotrectinib

[00184] 180mL of anhydrous DCM, compound1-2-Larotrectinib (80mmol, 42.91g),

and N,N-diisopropylethylamine (320mmol, 41.36g) were dosed in a 500 mL reaction flask;

2-(trimethylsilyl)ethoxymethyl chloride (SEM-Cl, 240mmol, 40.01g) was dosed in the

reaction mixture mixture at 25 °C; the obtained mixture was stirred at 25 °C to react

completely under the control of TLC; the reaction solution was washed once with

ammonium chloride solution (100mL, containing 37.2g of ammonium chloride), washed

twice with saturated sodium bicarbonate, 100ml each time, and washed once with brine

(100ml); the organic phase was dried over anhydrous magnesium sulfate and filtered; the

mixture was concentrated under reduced pressure; and the residue was subjected to silica

gel column chromatography (petroleum ether: ethyl acetate=5:1), thus obtaining the

product I-SEM-2-Larotrectinib (86.8%, 46.28g).

[00185] Example 34

[00186] Preparation of I-SEM-5-Larotrectinib by 2-(trimethylsilyl)ethoxymethyl

chloride etherification

[00187] The preparation method was the same as that in Example 33. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-SEM-5-Larotrectinib as a white solid with a yield of 86% (68.8mmol, 46g).

[00188] Example 35

[00189] Preparation of I-THP-2-Larotrectinib by tetrahydropyran etherification

F F O

7N -N OH :7N'-N

N N OH0 N N6

11 2 0

C 21H 22FIN60 2: 536.35 C 26H 30FING03:620.47 I - 2-Larotrectinib I -THP-2-Larotrectinib

[00190] 150 mL of DCM, compound 1-2-Larotrectinib (80mmol, 42.91g) and

pyridine p-toluenesulfonate (0.40g, 1.56mmol) were dosed in a 500 ml reaction flask and

stirred for 5 minutes; 3, 4-Dihydro-2H-pyran (13.66g, 168.46mmol) was dropwise dosed

and dissolved in the DCM (50ml) solution, and the solution was stirred overnight at room

temperature and then washed three times with brine (80gx3); the organic phase was dried

over anhydrous sodium sulfate and filtered with suction, the filtrate was concentrated to be

viscous, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 5:1), thus obtaining the product I-THP-2-Larotrectinib (98.6%,

48.94g).

[00191] Example 36

[00192] Preparation of I-THP-5-Larotrectinib by tetrahydropyran etherification

[00193] The preparation method was the same as that in Example 35. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-THP-5-Larotrectinib as a white solid with a yield of 98% (78.4mmol - 48g).

[00194] Example 37

[00195] Preparation of I-EE-2-Larotrectinib by 1-ethoxyethanol acetate (EE)

etherification

F F

N OH N'N KO NN ON N6 HN O OOII0HNN

C 2 1H 2 2 FIN 60 2 :536.35 C 25H 30FIN 60 3:608.46 I - 2-Larotrectinib I -EE-2-Larotrectinib

[00196] 150 mL of DCM, compound 1-2-Larotrectinib (80mmol, 42.91g), pyridine

p-toluenesulfonate (0.40g, 1.56mmol) and 1-ethoxyethanol acetate (C6H1203=132.16,

1.5e, 120mmol, 15.86g) were dosed in a 500 ml reaction flask and stirred for 1.5 hours at

room temperature; the solution was washed three times with brine (80gx3); the organic

concentrated, and the residue was subjected to silica gel column chromatography

(petroleum ether: ethyl acetate = 5:1), thus obtaining the product I-EE-2-Larotrectinib

(85.7%, 41.72g).

[00197] Example 38

[00198] Preparation of I-EE-5-Larotrectinib by 1-ethoxyethanol acetate (EE) etherification

[00199] The preparation method was the same as that in Example 37. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-EE-5-Larotrectinib as a white solid with a yield of 85% (68mmol 41g).

[00200] Example 39

[00201] Preparation of I-Als-2-Larotrectinib by allyl sulfonyl chloride etherification

F F N OH N' N O

NJ N 3~4NrN N ON-,

C21H22FIN6O2: 536.35 C24H26 FIN6O 4 S:640.47 I - 2-Larotrectinib I - Als-2-Larotrectinib

[00202] 180mL of DCM and compound 1-2-Larotrectinib (80mmol, 42.91g) were

dosed in a 500 ml reaction flask and then cooled to -30 °C, and allylsulfonyl chloride Als

(C3H5ClO2S=140.58, 2.5eq, 200mmol, 28.12g) was then dosed to react at this

temperature for 2 hours; the reaction mixture was heated to room temperature and stirred

to completely react for about 6 hours under the control of TLC; the reaction mixture was

concentrated under reduced pressure, the residue was dissolved in ethyl acetate (120 mL),

the mixture was washed twice with saturated sodium bicarbonate solution, 80mL each

time and then washed once with brine (80 mL); the organic phase was dried over

anhydrous magnesium sulfate, and filtered; the mixture was concentrated under reduced

pressure, and the residue was subjected to silica gel column chromatography (petroleum

ether: ethyl acetate=5:1), thus obtaining the product I-Als-2-Larotrectinib (86%, 44.06g).

[00203] Example 40

[00204] Preparation of I-Als-5-Larotrectinib by allyl sulfonyl chloride etherification

[00205] The preparation method was the same as that in Example 39. The compound -5-Larotrectinib was used as the raw material to obtain the product

I-Als-5-Larotrectinib as a white solid with a yield of 86% (68.8mmol 44g).

[00206] Example 41

[00207] Preparation of I-PMP-2-Larotrectinib by P-methoxyphenol etherification

(PMP-(p-Methoxyphenyl)ether)

F F OMe

PNN OH .-N Oc

N IN N 14N N., Nr

C 21H 22FIN 602 : 536.35 C28 H2 8FINS0 3:642.47 I - 2-Larotrectinib I -PMP-2-Larotrectinib

[00208] 100mL of THF, compound 1-2-Larotrectinib (80mmol, 42.91g),

triphenylphosphonium (1.3eq, 104mmol, 31.65g), diethyl azodicarboxylate (1.3eq,

104mmol, 18.11g), and p-methoxyphenol (3eq, 240mmol, 29.79g) were dosed in a 500 ml

reaction flask; the reaction mixture was heated to 80 °C, and stirred to completely react for

about 2 hours under the control of TLC; the mixture was concentrated under reduced

ether: ethyl acetate=5:1), thus obtaining the product I-PMP-2-Larotrectinib (81.3%,

41.6g).

[00209] Example 42

[00210] Preparation of I-PMP-5-Larotrectinib by P-methoxyphenol etherification

(PMP-(p-Methoxyphenyl)ether)

[00211] The preparation method was the same as that in Example 41. The

compound -5-Larotrectinib was used as the raw material to obtain the product

I-PMP-5-Larotrectinib as a white solid with a yield of 81% (64.8mmol, 43.6g).

[00212] Step 2: Preparation of labeled precursor

[00213] Example 43

[00214] Preparation of labeled precursor I(III)-SPIAd-Bz-2-Larotrectinib

F F

N- N OOCCH, F 3 CCOO N' N OOCCH,

N N FCCOO N N

0 0 C 28H 26FIN 60 3:640.46 C32 H 28F 71N0 7:868.50

I - Bz-2-Larotrectinib I -TfAc- Bz-2-Larotrectinib

F F

N -N O -N OOCC 6 H5 OOCCH, F3 CCOO> H I N FCCOO N N HNN Hil H O N N;H-( N 00

0 "H 0

C 2H 2 FylN 60 7:868.50 H C41H 42FIN 0 7:876.72 I-TfAc- Bz-2-Larotrectinib |(Ill)-SPIAd - Bz-2-Larotrectinib

[00215] 0.39mL of trifluoroacetic acid and 0.13mL of chloroform were dosed in a

mL reaction flask; urea-hydrogen peroxide complex (CO(NH2)2-H202 = 94.07,

1.36g,14.5mmol) and I-Bz-2-Larotrectinib (70.45mg, 0.11mmol) were then ndosed in the

mixed solution in a stirring way; the mixture was stirred at room temperature for 60

minutes and concentrated in vacuum to remove the solvent; 0.8mL of ethanol and 10%

sodium carbonate solution (0.5mL) containing SPIAd (25.3mg, 0.11mmol) were

sequentially dosed in the residue, and then the pH of the mixed solution was adjusted to 9

with 10% sodium carbonate solution (0.3mL); the mixture was stirred at room temperature

to react 70 minutes, and then the reaction solution was diluted with 5mL of water and

extracted three times with DCM, 5mL each time; the organic layers were combined, dried

over anhydrous magnesium sulfate, and concentrated in vacuum, and the the residue was

subjected to silica gel column chromatography (washed with ethyl acetate solution

containing 0-10% of methanol), thus obtaining the labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib as a white solid (18.32mg, 19%).

[00216] Example 44

[00217] I(III)-SPIAd-Bz-5-Larotrectinib was prepared by a method similar to the

method in Example 43, and the white solidI(III)-SPIAd-Bz-5-Larotrectinib was obtained

with a yield of 19% (18mmol, 18mg).

[00218] Example 45

[00219] Preparation of labeled precursor I(III)-SPIAd-Piv-2-Larotrectinib

F F 0 0

_N _N

0 N. 0 |- Piv-2-Larotrectinib I -Piv-TfAc-2-Larotrectinib

3CO N N6-aN N

F3CCOOOF H N

C 30H 30FylN 6 y: 846.50 H C 3 H FINO7 : 854.72 -Piv-TfAc-2-Larotrctinib I(II)-SPIAd-Pv-2-Larotrectinib

[00220] Trifluoroacetic anhydride (C4F603 = 210.04, 1.511 Cg/mL, 0.068mL,

0.489mmol, 102.75mg) and trifluoroacetic acid (CF3COOH=114.02, 1.5351g/cm3,

0.18mL, 0.276g, 2.43mmol) were dosed ina1mLround-bottomed flask; urea-hydrogen

peroxide (1.e25mmol, 106mg) was then slowly dosed in astirring way to react and release

heat and the temperature of the system was controlled below 30 °C; then,

I-Piv-2-Larotrectinib (155.12mg, 0.2mmol) was dosed, the reaction mixture was cooled

to about00 °C inan ice-water bath, andthenanhydrous sodiumsulfate(41mg, 0.5mmol)

was slowly dosed in the mixture; then, the reaction mixture was heated to 40°Cfor 1hour

to 6hours, and after TLC detected that the raw material I-Piv-2-Larotrectinib was

completely reacted, the reaction solution was diluted with 2.5mL of water and extracted

three times with DCM,S5mLeach time; theorganic phaseswere combined, dried over

anhydrous magnesium sulfate, filtered and concentrated under reduced pressure; the residue was subjected to silica gel column chromatography (eluted with ethyl acetate solution containing 0-10% of methanol), thus obtaining intermediate

I-Piv-TfAc-2-Larotrectinib as a white solid (143.91mg, 68%).

[00221] Na2CO3 (98.58mg, 0.93mmol) and MeCN (0.52ml), SPIAd (39.1mg,

0.17mmol) were dosed in a 10mL round-bottomed flask, and the heterogeneous mixture

was cooled to 0 °C in an ice-water bath and stirred vigorously for 5 minutes until the

mixture becomes milky white suspension. Intermediate I-Piv-TfAc-2-Larotrectinib

(143.91mg, 0.17 mmol) was dosed in the above vigorously stirred mixture at once, the

obtained mixture was continuously stirred at 0 °C to react for 2 hours, and the reaction

mixture gradually became a thick cream mixture. 10 mL of water was dosed, and the

obtained mixture was further stirred for 1 minute and then filtered to remove the fluffy

white suspended matter out of the beige biphase solution, the filter cake was washed twice

with water , 5 mL each time, the water needed to be vacuum dried every time the filter

cake was washed; the filter cake was then washed once with ether (10mL) and dried under

high vacuum, thus obtaining the labeled precursor I(III)-SPIAd-Piv-2-Larotrectinib as a

white solid (113.33mg, 78%, the total yield of 53% in two steps), with the m.p.of 100 °C

(decomposed).

[00222] Example 46

[00223] I(III)-SPIAd-Piv-5-Larotrectinib was prepared by a method similar to the

method in Example 45, and the white solid I(III)-SPIAd-Piv-5-Larotrectinib was obtained

with a yield of 78% (113mg - the total yield of 53% in two steps).

[00224] Example 47

[00225] Preparation of labeled precursor I(III)-SPIAd-Ac-2-Larotrectinib

F F 0 0 / - -N OAC oCH / : -0 .. NIN 3 F3CCOO N N§ H3

0 0 C 23H 24FING0 3:578.39 C 27H2 4F 7 N 6 07:804.42 1 - Ac-2-Larotrectinib I - Ac-TfAc-2-Larotrectinib

F F 00

/0'N'/N H N'N OC

F CCCOO N N 3 N H

C27H 24F 7 1NO07: 804.42 H C 36H 38FIN 6O 7:812.64 I -Ac-TfAc-2-Larotrectinib I(III)-SPIAd -Ac -2- Larotrectinib

[00226] Trifluoroacetic anhydride (0.068mL, 0.489mmol, 102.75mg),

trifluoroacetic acid (0.18mL, 0.276g, 2.43mmol), and 0.18mL of chloroform were dosed

in a 2mL round-bottomed flask; Oxone(136.36mg,0.225mmol) was then slowly dosed in a

stirring way; then, I-Ac-2-Larotrectinib (127.25mg - 0.22mmol) was dosed, the reaction

mixture was cooled to about 0 °C in an ice-water bath, and then anhydrous magnesium

sulfate (60mg, 0.5mmol) was slowly dosed in the mixture; then, the reaction mixture

reacted at room temperature of 25 °C for 6 hours, and after TLC detected that the raw

material I-Ac-2-Larotrectinib was completely reacted, the reaction solution was diluted

with 3mL of ice water and extracted three times with DCM, 5mL each time; the organic

phases were combined and washed three times with saline solution, 2mL each time; the

organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated

under reduced pressure; the residue was subjected to silica gel column chromatography

(eluted with ethyl acetate solution containing 0-10% of methanol), thus obtaining

intermediate I-Ac-TfAc-2-Larotrectinib as a light yellow solid (114.68mg - 0.143mmol,

64.8%).

[00227] NaHCO3 (78.12mg, 0.93mmol), acetone (0.52ml), and SPIAd (39.1mg,

0.17mmol) were dosed in a 2mL round-bottomed flask, and the heterogeneous mixture was cooled to 0 °C in an ice-water bath and stirred vigorously for 5 minutes until the mixture becomes milky white suspension. Intermediate I-Ac-TfAc-2-Larotrectinib

(143.91mg - 0.17mmol) was dosed in the above vigorously stirred mixture at once, the

mixture gradually became yellow; TLC tracked the reaction until the raw material was

completely converted; the insoluble matter was filtered off, and the reaction flask was

washed three times with dichloromethane, 0.12 mL each time, the insoluble matter was

rinsed with the washing liquid, and the yellow dichloromethane liquid was collected. The

collected liquid was concentrated under reduced pressure (at 20 °C) evaporated to remove

dichloromethane, and then dried for 1 hour under high vacuum, thus obtaining the labeled

precursor I(III)-SPIAd-Ac-2-Larotrectinib as a yellow solid (138.15mg, 46.6%).

[00228] Example 48

[00229] I(III)-SPIAd-Ac-5-Larotrectinib was prepared by a method similar to the

method in Example 47, and the white solid I(III)-SPIAd-Ac-5-Larotrectinib was obtained

with a yield of 46% (138mmol).

[00230] Example 49

[00231] Preparation of labeled precursor I(III)-SPIAd-Tf-2-Larotrectinib

F F 0 0

/ ~ 5 -N ~ ONC-N 'CF 3 F3CCOO-1 OACF /

N 6N F 3 CCOO N N N

0 0 C 23H 21F 4 1NO03:632.36 C 27H 23CIFIN 6 07: 838.86 S- Tf-2-Larotrectinib I - Tf-TfAc-2-Larotrectinib

F F

/0 N0 0F N.,~N OA" C ON.N OA.C

FaCCOO F3 CCOO F3 H,,H ON _"O HNIIIYN

C27H 21F 1OINGO: 858.39 H C36H 35F4IN 607:866.61 I -Tf-TfAc-2-Larotrectinib |(III)-SPIAd -Tf-2-Larotrectinib

[00232] Trifluoroacetic anhydride (0.068mL, 0.489mmol, 102.75mg),

stirring way; then, I-Tf-2-Larotrectinib (139.12mg - 0.22mmol) was dosed, the reaction

material I-Tf-2-Larotrectinib was completely reacted, the reaction solution was diluted

phases were combined and washed three times with brine, 2mL each time; the organic

phase was dried over anhydrous magnesium sulfate, filtered and concentrated under

reduced pressure; the residue was subjected to silica gel column chromatography (eluted

with ethyl acetate solution containing 0-10% of methanol), thus obtaining intermediate

I-Tf-TfAc-2-Larotrectinib as a light yellow solid (117.46mg - 0.137mmol, 62.2%).

[00233] K2C03 (128.34mg, 0.93mmol), methanol (0.52ml), and SPIAd (39.1mg,

0.17mmol) were dosed in a 2mL round-bottomed flask, and the heterogeneous mixture

mixture becomes milky white suspension. Intermediate I-Tf-TfAc-2-Larotrectinib

(117.46mg - 0.137mmol) was dosed in the above vigorously stirred mixture at once, the

obtained mixture was continuously stirred at 0 °C to react for 2 hours, and the reaction mixture gradually became yellow; TLC tracked the reaction until the raw material was completely converted; the insoluble matter was filtered off, and the reaction flask was washed three times with dichloromethane, 0.12 mL each time, the insoluble matter was rinsed with the washing liquid, and the yellow dichloromethane liquid was collected. The collected liquid was concentrated under reduced pressure (at 20 °C) evaporated to remove dichloromethane, and then dried for 1 hour under high vacuum, thus obtaining the labeled precursor I(III)-SPIAd-Tf-2-Larotrectinib as a yellow solid (53.9mg, 45.4%).

[00234] Example 50

[00235] I(III)-SPIAd-Tf-5-Larotrectinib was prepared by a method similar to the

method in Example 49, and the white solid I(III)-SPIAd-Tf-5-Larotrectinib was obtained

with a yield of 46% (54mg).

[00236] Example 51

[00237] Preparation of labeled precursor I(III)-SPIAd-CAc-2-Larotrectinib

F F

-N OOCCH 2CI -N OOCCH 2CI N ~F 3 CCOO-1 ~ N N F 3 CCOO N N

O O C 23H2 3CIFIN0 3: 612.83 C 27H 23CIFIN 07: 838.86 1 -CAc-2-Larotrectinib I - CAc-TfAc-2-Larotrectinib

F F

0 N'N OOCCH 2CI F3CCOO- 1 N'N I OOCCH 2CI H\ F3CCOO N N6 H HO N6 H, 0 0

C27H 23CIFINGO 7: 838.86 H C 36H37CIFIN 0 7:847.08 I - CAc-TfAc-2-Larotrectinib |(Ill)-SPIAd -CAc - 2-Larotrectinib

[00238] 0.3mL of trifluoroacetic acid, trifluoroacetic anhydride (0.9mL), oxone

(H3K5018S4 = 614.76, 101.44mg, 0.165mmol), and 0.13mL of chloroform were dosed in

a 2mL reactor in a stirring way, the obtained solution was stirred vigorously at room

temperature, and I-CAc-2-Larotrectinib (67.41mg, 0.11mmol) was then dosed in the uniform solution; the obtained mixture was cooled to 0 °C; anhydrous sodium acetate

(0.53g, 6.45mmol) was slowly dosed in the vigorously stirred mixture; then, the mixture

was heated to 40 °C and vigorously stirred to react for 2 hours, and TLC tracked the

reaction until the raw material was completely converted; the mixture was cooled to room

temperature, diluted with 2 mL of water, and extracted three times with dichloromethane,

3ml each time; the organic phases were combined and extracted, then washed three times

with saturated brine, 3ml each time, dried with anhydrous sodium sulfate, filtered, and

concentrated; the residue was washed with n-heptane/ethyl acetate and filtered, thus

obtaining a yellow solid I-CAc-TfAc -2-Larotrectinib (57.12mg, 0.068mmol, 61.9%).

[00239] KHC03 (93mg, 0.93mmol), ethanol (0.52ml), and SPIAd (39.1mg,

0.l7mmol) were dosed in a 2mL reactor, and the heterogeneous mixture was cooled to 0

°C in an ice-water bath and stirred vigorously for 5 minutes until the mixture becomes

milky white suspension. Intermediate I-CAc-TfAc-2-Larotrectinib (57.12mg, 0.068 mmol)

was dosed in the above vigorously stirred mixture at once, the obtained mixture was

continuously stirred at 0 °C to react for 2 hours, and the reaction mixture gradually

became yellow; TLC tracked the reaction until the raw material was completely converted;

the insoluble matter was filtered off, and the reaction flask was washed three times with

dichloromethane, 0.12 mL each time, the insoluble matter was rinsed with the washing

liquid, and the yellow dichloromethane liquid was collected. The collected liquid was

concentrated under reduced pressure (at 20 °C) evaporated to remove dichloromethane,

and then dried for 1 hour under high vacuum, thus obtaining the labeled precursor

I(III)-SPIAd-CAc-2-Larotrectinib as a yellow solid (26.84mg, 46.6%).

[00240] Example 52

[00241] I(III)-SPIAd-CAc-5-Larotrectinib was prepared by a method similar to the

method in Example 51, and the white solid I(III)-SPIAd-CAc-5-Larotrectinib was obtained with a yield of 46% (138mmol).

[00242] Example 53

[00243] Preparation of labeled precursor I(III)-SPIAd-DCAc-2-Larotrectinib

F F

N'N OOCCHC1 2 N-N OOCCHC1 2 F 3 CCOO. N F3CCOO NN

C 23H 22 Cl 2FIN 60 3: 647.27 C 27 H 2 2C12FylN 6Oy: 873.30 1 - DCAc-2-Larotrectinib I - DCAc-TfAc-2-Larotrectinib

F F

N'-N OOCCHC1 2 H N N OOCCHCl 2 F Ox

F 3CCOO N 7JNI 0 0 0 HO N N6 N N Hh

C27H 22C1 2FyN 60 7:873.30 C36H 36Cl2FIN 60 7:881.52 1 - DCAc-TfAc-2-Larotrectinib H I(Ill)-SPIAd - DCAc-2-Larotrectinib

[00244] 0.3mL of trifluoroacetic acid, trifluoroacetic anhydride (0.9mL), oxone

(H3K50184= 614.76, 101.44mg 0.165mmol), and 0.l3mL of chloroform were dosed in a

2mL reactor in a stirring way, the obtained solution was stirred vigorously at room

temperature, and I-DCAc-2-Larotrectinib(71.2mg - 0.11mmol) was then dosed in the

uniform solution; the obtained mixture was cooled to 0 °C; anhydrous sodium acetate

concentrated; the residue was washed with N-heptane ethyl acetate and filtered, thus

obtaining a yellow solid I-DCAc-TfAc-2-Larotrectinib (65.32mg, 0.075mmol, 68%).

[00245] 10% sodium carbonate aqueous solution (w/v.0.75mL, 0.33M) containing

auxiliary acid adamantane (SPI-Adaman, O.1mmol, 23mg) and ethanol (1mL) were dosed

in a 2mL reaction flask, and then I -DCAc-TfAc-2-Larotrectinib (0.096mmol, 84.53mg)

was dosed quickly; the reaction mixture was stirred vigorously for 0.5 hour at room

temperature, and TLC tracked the reaction until the raw material was completely

converted; the reaction mixture was diluted with 2mL of water, and extracted three times

with dichloromethane, 3ml each time; the organic extract phases were combined, dried

with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure; the

residue was subjected to silica gel column chromatography and eluted with

n-heptane/ethyl acetate=10/1, thus obtaining the labeled precursor

I(III)-SPIAd-DCAc-2-Larotrectinib as a white solid (43.16mg, 46.8%, the total yield of

46.8% in two steps).

[00246] Example 54

[00247] I(III)-SPIAd-DCAc-5-Larotrectinib was prepared by a method similar to

the method in Example 53, and the white solid I(III)-SPIAd-DCAc-5-Larotrectinib was

obtained with a yield of 78% (43mg 47%, the total yield of 46.8% in two steps).

[00248] Example 55

[00249] Preparation of labeled precursor I(III)-SPIAd-Moc-2-Larotrectinib

F F

N N NN 0FCCOO

C 23H 24 FIN 604:594.39 C2 7H 4 0FylN 6 0:842.66 I- Moc-2-Larotrectinib I-Moc-TfAc-2-Larotrectinib

F F 0 0

F 3 CCOONN OAO ON 0 F3CCOO N NH N N6

C27H4FylN6O 8:842.66 H C36H 38FINSO0: 828.64 1 -Moc-TfAc-2-Larotrectinib |(III)-SPIAd -Moc -2-Larotrectinib

[00250] 0.36 mL of trifluoroacetic acid and 0.18 mL of chloroform were dosed in a

2 mL reactor; I-Moc-2-Larotrectinib (89.16mg, 0.15mmol) and oxone (110.66mg,

0.18mmol) were dosed in a stirring way; the mixture was heated to 45 °C and stirred to

react for 3 hours; then, the reaction mixture was concentrated under reduced pressure to

remove the solvent; 1.2 mL of ethanol and 0.68 mL of SPIAd (34.5 mg, 0.15 mmol)

dissolved in 10% sodium carbonate solution were dosed in the residue and the pH of the

mixture was adjusted to 12 with the 10% sodium carbonate solution. The mixture was

stirred at 50°C for 3 hours, and TLC tracked the reaction until the raw material was

completely converted; the reactant was diluted with 3mL of water, and extracted three

times with DCM, 5 mL each time; the organic phases were combined, dried over

magnesium sulfate, and concentrated under reduced pressure; and the residue was

subjected to silica gel column chromatography (eluted with 0-40% ethyl acetate-petroleum

ether solution), thus obtaining the labeled precursor I(III)-SPIAd-Moc-2-Larotrectinib as a

white solid (38.91mg, 31.3%).

[00251] Example 56

[00252] I(III)-SPIAd-Moc-5-Larotrectinib was prepared by a method similar to the

method in Example 55, and the white solid I(III)-SPIAd-Moc-5-Larotrectinib was

obtained with a yield of 31% (39mg).

[00253] Example 57

[00254] Preparation of labeled precursor I(III)-SPIAd-Eoc-2-Larotrectinib

F F 0 O,\ 0

N'N OAO -N O AO - ! F 3CCOO- 1 N HN F 3 CCOO NI N

0 0

C 24H 26FIN 604 :608.41 C 28H 2 6F 71N OS: 6 834.44 1 - Eoc-2-Larotrectinib I - Eoc-TfAc-2-Larotrectinib

F F 0 0

N'N O-/ O N'N O AO F 3CCOO- N H . H NIN F3CCOO N H,"'HIO

C28H 26F 71N O0: 6 834.44 H C 37H 40FING0 8: 842.66 |-Eoc-TfAc-2-Larotrectinib |(III)-SPIAd-Eoc-2-Larotrectinib

[00255] 0.36 mL of trifluoroacetic acid and 0.18 mL of chloroform were dosed in a

2 mL reactor; I-Eoc-2-Larotrectinib (91.26mg 0.15 mmol) and

Oxone(110.66mg,0.8mmol) were dosed in a stirring way; the mixture was heated to 37

°C and stirred to react for 3 hours; then, the reaction mixture was concentrated under

reduced pressure to remove the solvent; 1.2 mL of ethanol and 0.68 mL of SPIAd (34.5

mg, 0.15 mmol) dissolved in 10% sodium carbonate solution were dosed in the residue,

and the pH of the mixture was adjusted to 11 with the 10% sodium carbonate solution. The

mixture was stirred at 60°C for 2 hours and TLC tracked the reaction until the raw

material was completely converted; the reactant was diluted with 3mL of water and

extracted three times with DCM, 5 mL each time; the organic phases were combined,

dried over magnesium sulfate, and concentrated under reduced pressure; and the residue

was subjected to silica gel column chromatography (eluted with 0 -4 0 % ethyl

acetate-petroleum ether solution), thus obtaining the labeled precursor

I(III)-SPIAd-Eoc-2-Larotrectinib as a white solid (33.57mg, 33.2%).

[00256] Example 58

[00257] I(III)-SPIAd-Eoc-5-Larotrectinib was prepared by a method similar to the

method in Example 57, and the white solid I(III)-SPIAd-Eoc-5-Larotrectinib was obtained with a yield of 33% (34mg).

[00258] Example 59

[00259] Preparation of labeled precursor I(III)-SPIAd-Boc-2-Larotrectinib

F F

N OBoc O N'N OBoc HI

N N H N N N

C 26H 30FIN604: 636.47 H C 39H 44FIN080: 870.72 I - Boc-2-Larotrectinib I(III)-SPIAd - Boc-2-Larotrectinib

[00260] 0.8mL of dichloromethane was dosed in a 2mL reactor, and

I-Boc-2-Larotrectinib (89.11mg, 0.14mmol) and mCPBA (40mg, 0.18mmol) were then

dosed in a stirring way; the mixture was stirred vigorously at room temperature and

reacted for 1 hour to 2 hours, TLC tracked the reaction until the raw material was

completely reacted, and the obtained material was for later use.

[00261] KOH = 56.1 (47.12mg, 0.84mmol), dichloromethane (0.47ml), and SPIAd

(39.1mg, 0.17mmol) were dosed in a 2mL reactor, and the heterogeneous mixture was

cooled to 0 °C in an ice-water bath and stirred vigorously for 5 minutes until the mixture

becomes milky white suspension (pH12). The material obtained above for later use was

dosed in the above vigorously stirred mixture at once, the obtained mixture was

I(III)-SPIAd-Boc-2-Larotrectinib as a yellow solid (49.74mg, 40.8%) with the m.p.of 88.6

°C (decomposed).

[00262] Example 60

[00263] I(III)-SPIAd-Boc-5-Larotrectinib was prepared by a method similar to the

method in Example 59, and the white solid I(III)-SPIAd-Boc-5-Larotrectinib was obtained

with a yield of 40% (49mg).

[00264] Example 61

[00265] Preparation of labeled precursor I(III)-SPIAd-Troc-2-Larotrectinib

F F -O CCla N'N O CCl

0N F3CCOO N N O

N HN-(N 3COH-,6

0 0 C24 H 23C1 3FIN 604:711.74 C 2 H2 3C 3F7 1N 608 :937.77 I -Troc-2-Larotrectinib I - Troc-TfAc-2-Larotrectinib

F

F OGCla O N O CCl3 Fa\CC 3 N H N O F 3 CCOO-I0 F 3CCOO' /N N N6ll 1N N HN- H

C28H23CI3 F7 IN6Og:937.77 H C37 H 37 Cl3FIN 6 09:945.99 I - Troc-TfAc-2-Larotrectinib |(III)-SPAd -Troc-2-Larotroctinib

[00266] 0.36 mL of trifluoroacetic acid and 0.18 mL of chloroform were dosed in a

2 mL reactor; I-Troc-2-Larotrectinib (106.76mg 0.15 mmol) and

Oxone(110.66mg,0.8mmol) were dosed in a stirring way; the mixture was heated to 70

°C and stirred to react for 2 hours; then, the reaction mixture was concentrated under

reduced pressure to remove the solvent; 1.2 mL of ethanol and 0.68mL of SPIAd (34.5mg,

0.15mmol) dissolved in 10% sodium carbonate solution were dosed in the residue, and the

pH of the mixture was adjusted to 9 with the 10% sodium carbonate solution. The mixture

was stirred at room temperature for 70 minutes; TLC tracked the reaction until the raw

extracted three times with DCM, 5 mL each time; the organic phases were combined, dried over magnesium sulfate, and concentrated under reduced pressure; and the residue was subjected to silica gel column chromatography (eluted with 0 -4 0 % ethyl acetate-petroleum ether solution), thus obtaining the labeled precursor

I(III)-SPIAd-Troc-2-Larotrectinib as a white solid (53.92mg, 38%).

[00267] Example 62

[00268] I(III)-SPIAd-Troc-5-Larotrectinib was prepared by a method similar to the

method in Example 61, and the white solid I(III)-SPIAd-Troc-5-Larotrectinib was

obtained with a yield of 38% (53mg).

[00269] Example 63

[00270] Preparation of labeled precursor I(III)-SPIAd-Teoc-2-Larotrectinib

F 0 F

-N, _N OAO

N N NJ FNCN 0 0N

C 27 H34 FIN 60 4 Si:680.59 C31 H 34 F7IN 60 8 Si:906.63 |-Teoc-2-Larotrectinib I -Teoc-TfAc-2-Larotrectinib F 3 CCOO - N i3'C<O HN_ Si ' HH, O O--S F F

\ ~ N'N 0 ~ N'N O

H N0 N HHOON N O '"'H C31H 36 IN60O F7 7 Si:892.64 H C 40H 48FN6 0 Si:914.85 I-Teoc-TfAc-2-Larorectinib I (I)-SPIAd -Teoc-2-Larotre ctinib

[00271] Trifluoroacetic anhydride (0.068mL, 0.489mmol, 102.75mg),

in a2mL round-bottomed flask; Oxone(136.36mg,0.225mmo) was then slowly dosed in a

stirring way; then, I-Teoc-2-Larotrectinib (147.73mg -0.22mmol) was dosed, the reaction

mixture was cooled to about00 °C inan ice-water bath, andthenanhydrous magnesium

sulfate (60mg, 0.5mmol) was slowly dosed in the mixture; then, the reaction mixture reacted at room temperature of 25 °C for 6 hours, and after TLC detected that the raw material I-Tf-2-Larotrectinib was completely reacted, the reaction solution was diluted with 3mL of ice water and extracted three times with DCM, 5mL each time; the organic phases were combined and washed three times with brine, 2mL each time; the organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure; the residue was subjected to silica gel column chromatography (eluted with ethyl acetate solution containing 0-10% of methanol), thus obtaining intermediate

I-teoc-TfAc-2-Larotrectinib as a light yellow solid (153.96mg, 0.173mmol, 78.4%).

[00272] KOH(52.08mg, 0.93mmol), MeCN(0.52ml) and SPIAd(39.1mg, 0.17mmol)

were dosed in a 10mL round-bottomed flask, and the heterogeneous mixture was cooled to

milky white suspension. Intermediate I-Teoc-TfAc-2-Larotrectinib (143.91mg, 0.17mmol)

became a thick cream mixture. 10 mL of water was dosed, and the obtained mixture was

further stirred for 1 minute and then filtered to remove the fluffy white suspended matter

out of the beige biphase solution, the filter cake was washed twice with water, 5mL each

time, the water needed to be vacuum dried every time the filter cake was washed; the filter

cake was then washed once with ether (10mL) and dried under high vacuum, thus

obtaining the labeled precursor I(III)-SPIAd-Teoc-2-Larotrectinib as a white solid

(121.31mg, 78%, the total yield of 61.1% in two steps), with the m.p.of 115 °C

(decomposed).

[00273] Example 64

[00274] I(III)-SPIAd-Teoc-5-Larotrectinib was prepared by a method similar to the

method in Example 63, and the white solid I(III)-SPIAd-Teoc-5-Larotrectinib was obtained (121mg, 78%, the total yield of 61% in two steps).

[00275] Example 65

[00276] Preparation of labeled precursor I(III)-SPIAd-TBS-2-Larotrectinib

F F

-N OTBS N~N OTBS - ~F 3CCOO.... N N FCCOO N N HN-, HN 0 0 C 27H 36FIN 60 2Si:650.61 C 31H 36F 71NG0 6Si:876.64 1 -TBS-2-Larotrectinib I - TBS-TfAc-2-Larotrectinib

F F

FCCOO ' -N'N OTBS 0 N.-N OTBS

F 3CCOO N N N 0 HN-W H,. ' 0 N HN N

0 'H 0 C 31H 36F 71N 6O 6Si: 876.64 H CwHSOFIN 6O6Si: 884.86 1 -TBS-TfAc-2-Larotrectinib |(Ill)-SPIAd -TBS - 2-Larotrectinib

[00277] 0.39mL of trifluoroacetic acid and 0.13mL of acetonitrile were dosed in a

mL reactor and the mixture was stirred and cooled to 0 °C; Oxone (100mg,0.165mmol)

and I-TBS-2-Larotrectinib (70.2mg, 0.11mmol) were sequentially dosed in the mixture;

then, the mixture was stirred at 0 °C to react for 30 minutes; 0.8mL of ethanol and 0.5mL

of SPIAd (25.3mg, 0.11mmol) dissolved in 10% sodium carbonate solution were dosed in

the residue, and the pH of the mixture was adjusted to 9 with 0.3mL of the 10% sodium

carbonate solution. The mixture was stirred at room temperature for 70 minutes and then

diluted with 5mL of water and extracted three times with DCM, 5mL each time; the

organic phases were combined, dried over magnesium sulfate, and concentrated under

reduced pressure; and the residue was subjected to silica gel column chromatography

(eluted with ethyl acetate solution containing 0-10% of methanol), thus obtaining the

labeled precursor I(III)-SPIAd-TBS-2-Larotrectinib as a white solid (17.52mg, 18%).

[00278] Example 66

[00279] I(III)-SPIAd-TBS-5-Larotrectinib was prepared by a method similar to the

method in Example 65, and the white solid I(III)-SPIAd-TBS-5-Larotrectinib was

obtained (38mg, 30%).

[00280] Example 67

[00281] Another preparation method of labeled precursor

I(III)-SPIAd-TBS-2-Larotrectinib

F F / N'N OTBS N- N OTBS

N N6H H N N

0 '"H 0 C 27H 36FIN 602Si:650.61 H C 4 0HS 0FIN 6 0GSi: 884.86

1 -TBS-2-Larotrectinib |(Ill)-SPIAd -TBS - 2-Larotrectinib

[00282] 0.8mL of dichloromethane was dosed in a 2mL reactor, and

I-TBS-2-Larotrectinib (89.11mg, 0.14mmol) and mCPBA (40mg, 0.18mmol) were then

completely reacted, and the obtained material was for later use.

[00283] KOH = 56.1 (47.12mg, 0.84mmol), dichloromethane (0.47ml), and SPIAd

dosed in the above vigorously stirred mixture at once, the obtained mixture was

dichloromethane, 0.12 mL each time, the insoluble matter was rinsed with the washing liquid, and the yellow dichloromethane liquid was collected. The collected liquid was concentrated under reduced pressure (at 20 °C) evaporated to remove dichloromethane, and then dried for 1 hour under high vacuum, thus obtaining the labeled precursor

I(III)-SPIAd-TBS-2-Larotrectinib as a yellow solid (38.16mg, 30.8% in total).

[00284] Example 68

[00285] I(III)-SPIAd-TBS-5-Larotrectinib was prepared by a method similar to the

method in Example 67, and the white solid I(III)-SPIAd-TBS-5-Larotrectinib was

obtained (38mg, 30%).

[00286] Example 69

[00287] Preparation of labeled precursor I(III)-SPIAd-TBDPS-2-Larotrectinib

F F

N' OTBDPS N'N OTBDPS I ~F 3CCOO-.

N N - F3 CCOO N N 0 0 C3 7H4 0FIN 602Si:774.75 C4 1H 40FyN 606Si: 1000.79 1 -TBDPS-2-Larotrectinib I - TBDPS-TfAc-2-Larotrectinib

F F

OTBDPS 0 NN OTBDPS 0\CCN'N F3CCOO N N, HON 0

O "'IH 0 C41H 40FylNSOSSi:1000.79 H C 50H4FIN 6 O 6Si:1009.01 1 - TBDPS-TfAc-2-Larotrectinib I(Ill)-SPIAd -TBDPS - 2-Larotrectinib

[00288] 0.53 mL of trifluoroacetic acid, 0.18mL of chloroform and magnesium

chloride (MgCl2, 0.95mg, 0.Olmmol) were dosed in a 2 mL reactor;

I-TBDPS-2-Larotrectinib (116.22mg, 0.15 mmol) and Oxone (H3K5018S4 = 614.76,

136.36mg, 0.225mmol) were dosed in a stirring way; the mixture was heated to 80 °C and

stirred to react for 3 hours, and TLC tracked the reaction until the raw material was

completely reacted; the reaction mixture was filtered to remove the solid and then

concentrated under reduced pressure to remove the solvent; 1.2mL of ethanol and 0.68 mL of SPIAd (34.5mg, 0.15mmol) dissolved in 10% sodium carbonate solution were dosed in the residue, and the pH of the mixture was adjusted to 9 with 0.41mL of the 10% sodium carbonate solution. The mixture was stirred at room temperature for 70 minutes; TLC tracked the reaction till the end of the reaction; the reactant was diluted with 3mL of water, and extracted three times with DCM, 5 mL each time; the organic phases were combined, dried over anhydrous magnesium sulfate, concentrated under reduced pressure; and the residue was subjected to silica gel column chromatography (eluted with 0-40% ethyl acetate-petroleum ether solution), thus obtaining the labeled precursor

I(III)-SPIAd-PMP-2-Larotrectinib as a white solid (52.97mg, 35%).

[00289] Example 70

[00290] I(III)-SPIAd-TBS-5-Larotrectinib was prepared by a method similar to the

method in Example 69, and the white solid I(III)-SPIAd-TBS-5-Larotrectinib was

obtained (53mg, 35%).

[00291] Example 71

[00292] Preparation of labeled precursor I(III)-SPIAd-TIPS-2-Larotrectinib

F F NN OTIPS / N'N OTIPS

KiiY~ N F HN_ Kjjj 3 CCOO N N

0 0 C30 H42FING0 2Si:692.69 C34H 42F7 N 6 O6Si: 918.72 1 - TIPS-2-Larotrectinib I - TIPS-TfAc-2-Larotrectinib

F F

-N OTIPS O N'N OTIPS F 3CCOO- HH~OTP 0 N HN_ N N F 3CCO N 0 ' HHN N

C 34H42F 7lN 6O0SI: 918.72 H C4 3H56FIN 6O 6Si: 926.94 1 -TIPS-TfAc-2-Larotrectinib |(Ill)-SPIAd -TIPS - 2-Larotrectinib

[00293] 4mL of dichloromethane was dosed in a 2mL reactor and trifluoroacetic

anhydride (0.16mL, 0.24g, 1.14mmol), Oxone (67.63mg, 0.11mmol), 2,2,6,6-tetramethyl piperidine-oxide (TEMPO, 0.001mmol, 0.056mg), and anhydrous lithium chloride (LiCl,

0.42mg, 0.01mmol) were then dosed in a stirring way; the mixture was stirred well at

room temperature, and I-TIPS-2-Larotrectinib (86.33mg, 0.11mmol) was then dosed; the

resulting mixture was reacted at room temperature for 2 hours, and TLC tracked the

reaction process; after the raw material disappeared, the solid was removed by filtration;

then, the organic phase was concentrated, and the residue was separated by silica gel

column chromatography (eluted with developing solvent: 0-50% ethyl acetate-n-heptane,

v/v); the target product eluate was collected and concentrated, thus obtaining a colorless

solid I-TIPS-TfAc-2-Larotrectinib (63.67mg, 0.069 mmol, 63%).

[00294] 10% sodium carbonate aqueous solution (w/v.0.75mL, 0.33M) containing

auxiliary acid adamantane (SPI-Adaman, 0.1mmol, 23mg) and ethanol (1mL) were dosed

in a 2mL reaction flask, and then I-TIPS-TfAc-2-Larotrectinib (0.069mmol, 63.67mg) was

dosed quickly; the reaction mixture was stirred vigorously for 4 hour at room temperature,

and TLC tracked the reaction until the raw material was completely converted; the reacant

was diluted with 3mL of water, and extracted three times with DCM, 5ml each time; the

organic phases were combined, dried with anhydrous sodium sulfate, filtered, and

concentrated under reduced pressure; the residue was subjected to silica gel column

chromatography (eluted with developing solvent: 0- 5 0 % ethyl acetate-petroleum ether,

v/v); the target product eluate was collected and concentrated, thus obtaining labeled

precursor I(III)-SPIAd-TIPS-2-Larotrectinib as a white solid (42.82mg, the total yield of

42% in two steps).

[00295] Example 72

[00296] I(III)-SPIAd-TIPS-5-Larotrectinib was prepared by a method similar to the

method in Example 71, and the white solid I(III)-SPIAd-TIPS-5-Larotrectinib was

obtained (42mg, the total yield of 42% in two steps).

[00297] Example 73

[00298] Preparation of labeled precursor I(III)-SPIAd-PMB-2-Larotrectinib

F OMe F r\, OMe a O~e

N N F 3 CCOO 1 N N O6

0 0 C 33 H3OF7 1N 6 07 :882.53 C29H 30FIN 60 3:656.50 I - PMB-TfAc-2-Larotrectinib I - PMB-2-Larotrectinib

F F

O OMe F3CCOO-. N-NOMe J H F3CCOO N H N N HN-_ , HHN_

C33H3F7IN67O,:882.53 H C 42 H 44FIN 6 0 7 :890.75 I - PMB-TfAc-2-Larotrectinib I(III)-SPIAd -PMB-2-Larotrectinib

[00299] 0.53mL of trifluoroacetic acid and 0.18mL of chloroform were dosed in a 2

mL reactor; I-PMB-2-Larotrectinib (98.47mg, 0.15 mmol), sodium chloride (0.59mg,

0.Olmmol), and Oxone (H3K5018S4 = 614.76, 138.32mg, 0.225mmol) were dosed in a

stirring way; the mixture was heated to 80 °C and stirred to react for 3 hours, and TLC

tracked the reaction until the raw material was completely reacted; the reaction mixture

was then concentrated under reduced pressure to remove the solvent; 1.2mL of ethanol

and 0.68 mL of SPIAd (34.5mg, 0.15mmol) dissolved in 10% sodium carbonate solution

were dosed in the residue, and the pH of the mixture was adjusted to 9 with 10% sodium

carbonate solution. The mixture was stirred at room temperature for 70 minutes; TLC

tracked the reaction till the end of the reaction; the reactant was diluted with 3mL of water,

and extracted three times with DCM, 5 mL each time; the organic phases were combined,

dried over anhydrous magnesium sulfate, concentrated under reduced pressure; and the

residue was subjected to silica gel column chromatography (eluted with 0-40% ethyl

acetate-petroleum ether solution), thus obtaining the labeled precursor

I(III)-SPIAd-PMB-2-Larotrectinib as a yellow solid (44.89mg, 33.6%).

[00300] Example 74

[00301] I(III)-SPIAd-PMB-5-Larotrectinib was prepared by a method similar to the method in Example 73, and the white solid I(III)-SPIAd-PMB-5-Larotrectinib was obtained (45mg, 34%).

[00302] Example 75

[00303] Preparation of labeled precursor I(III)-SPIAd-MOM-2-Larotrectinib

F F -N OMOM -N OMOM

CCOO N F3 N N HN--( N H

0 0

C 23 H26 FIN 6 03 :580.40 C 27 H26 F 7 1N 6 07 :806.43 I -MOM-2-Larotrectinib I - MOM-TfAc-2-Larotrectinib

OMOM O -N OMOM F CCOO 3 N N MH N N

C 27H2 6F 7IN6 0 7 :806.43 H C36H40 FIN6 0 7 :814.65 I - MOM-TfAc-2-Larotrectinib I(III)-SPIAd - MOM-2-Larotractinib

[00304] 4mL of acetonitrile was dosed in a 2mL reactor and Oxone (H3K5018S4=

614.76, 67.63mg, 0.11mmol) and 0.05mL of acetone were then dosed in a stirring way; the

mixture was stirred well at room temperature; I-MOM-2-Larotrectinib(63.84mg

0.11mmol) was then dosed; the resulting mixture was heated to 60 °C and reacted for 2

hours and the reaction solution was then cooled to room temperature; trifluoroacetic

anhydride (0.16mL, 0.24g, 1.14mmol) was dosed dropwise, the resulting mixture reacted

at room temperature for 2 hours, and TLC tracked the reaction process; after the raw

material disappeared, the solid was removed by filtration; the residue was separated by

silica gel column chromatography (eluted with developing solvent: 0-50% ethyl

acetate-n-heptane, v/v); the target product eluate was collected and concentrated, thus

obtaining a yellow solid I-MOM-TfAc-2-Larotrectinib (48.79mg, 0.061 mmol, 55%).

[00305] 10% sodium carbonate aqueous solution (w/v.0.75mL, 0.33M) containing auxiliary acid adamantane (SPI-Adaman,0.1mmol,23mg) and ethanol (1mL) were dosed in a 2mL reaction flask, and then I-MOM-TfAc-2-Larotrectinib (0.061mmol,48.79mg) was dosed quickly; the reaction mixture was stirred vigorously for 3 hour at room temperature, and TLC tracked the reaction until the raw material was completely converted; the reacant was diluted with 3mL of water, and extracted three times with dichloromethane, 5ml each time; the organic extract phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure; the residue was subjected to silica gel column chromatography (eluted with developing solvent:

-50% ethyl acetate-petroleum ether, v/v); the target product eluate was collected and

concentrated, thus obtaining labeled precursor I(III)-SPIAd-MOM-2-Larotrectinib as a

white solid (38.53mg, the total yield of 43% in two steps).

[00306] Example 76

[00307] I(III)-SPIAd-MOM-5-Larotrectinib was prepared by a method similar to

the method in Example 75, and the white solid I(III)-SPIAd-MOM-5-Larotrectinib was

obtained (38mg, the total yield of 43% in two steps).

[00308] Example 77

[00309] Preparation of labeled precursor I(III)-SPIAd-MEM-2-Larotrectinib

F F

N OMEM /.--N OMEM - F 3CCOO-I N H

N N FCCOO NNI

0 0 C25 H30 FIN 6 04 :624.46 C 29H 3 0F7 IN 6 08 :850.49 I -MEM-2-Larotrectinib I - MEM-TfAc-2-Larotrectinib

F F OMEM H N OMEM N-N

F 3CCOO N NH N N6

0 H 0 C 29 H3 OF7 IN 6 0 8 :850.49 H C3 gH 4 4FIN 6 Os:858.71 I - MEM-TfAc-2-Larotrectinib |(Ill)-SPIAd - MEM-2-Larotrectinib

[00310] 0.4mL of acetonitrile was dosed in a 2mL reactor and Oxone (67.63mg

0.11mmol) and NaBr(0.011mmol,1.13mg) were then dosed in a stirring way; the mixture

was stirred well at room temperature; I-MEM-2-Larotrectinib(68.69mg - 0.11mmol) was

then dosed; the resulting mixture was heated to 60 °C and reacted for 2 hours and the

reaction solution was then cooled to room temperature; trifluoroacetic anhydride (0.16mL,

0.24g, 1.14mmol, 1.511 C g/mL) was dosed dropwise, the resulting mixture reacted at

room temperature for 2 hours, and TLC tracked the reaction process; after the raw material

disappeared, the solid was removed by filtration; the organic phase was concentrated and

the residue was then separated by silica gel column chromatography (eluted with

developing solvent: 0-50% ethyl acetate-n-heptane, v/v); the target product eluate was

collected and concentrated, thus obtaining a colorless solid I-MEM-TfAc-2-Larotrectinib

(52.39mg, 0.062mmol, 56%).

[00311] 10% sodium carbonate aqueous solution (w/v.0.75mL, 0.33M) containing

in a 2mL reaction flask, and then I-MEM-TfAc-2-Larotrectinib (0.062mmol, 52.39mg)

was dosed quickly; the reaction mixture was stirred vigorously for 4 hour at room

temperature, and TLC tracked the reaction until the raw material was completely

converted; the reaction mixture was diluted with 3mL of water, and extracted three times

with dichloromethane, 5ml each time; the organic extract phases were combined, dried

with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure; the residue was subjected to silica gel column chromatography (eluted with developing solvent: 0-50% ethyl acetate-petroleum ether, v/v); the target product eluate was collected and concentrated, thus obtaining labeled precursor I(III)-SPIAd-MEM-2-Larotrectinib as a white solid (30.23mg, the total yield of 32% in two steps).

[00312] Example 78

[00313] I(III)-SPIAd-MEM-5-Larotrectinib was prepared by a method similar to

the method in Example 77, and the white solid I(III)-SPIAd-MEM-5-Larotrectinib was

obtained (30mg, the total yield of 32% in two steps).

[00314] Example 79

[00315] Preparation of labeled precursor I(III)-SPIAd-SEM-2-Larotrectinib

F F

N 0 0 N OSEM - F 3CCOO-

N N S F 3CCOO KJN N

0 0 C2 7 H36 FIN 6 03 Si:666.61 C31 H36 F7 IN 60 7 Si:892.64 I -SEM-2-Larotrectinib I - SEM-TfAc-2-Larotrectinib

OSEM O N'N OSEM N

F 3CCOO N NH I N N

C 3 1H 36 F7 IN 6 07 Si:892.64 H C 40 HSOFIN 6 0 7 Si:900.86 I - SEM-TfAc-2-Larotrectinib |(Ill)-SPIAd - SEM-2-Larotrectinib

[00316] 0.13mL of acetonitrile, trifluoroacetic acid (0.39mL, 5.26mmol, 0.6g,

1.5351g/cm3), and Oxone (100mg, 0.165mmol) were dosed in a 2mL reactor; the mixture

was cooled to 0 °C to 5 °C in an ice-water bath and stirred to react for 0.5 hour;

I-SEM-2-Larotrectinib (73.32mg, 0.11mmol) was slowly dosed dropwise to the vigorously

stirred mixture; the mixture was stirred and reacted at 0 to 5 °C for 1 hour, and the reaction

solution was diluted with ethanol (0.5mL); 10% sodium carbonate solution (0.1mL)

containing auxiliary acid adamantane (SPI-Adaman, 25.3mg, 0.11mmol) was then dosed quickly, and the pH of the mixed solution was adjusted to about 9 with 10% sodium carbonate solution (note: bubbles were generated in this step). The mixture was stirred at 0

°C to 5 °C for 3 hours and then diluted with 5mL of water, and extracted three times with

DCM, 5mL each time; the organic phases were combined, dried over magnesium sulfate,

and concentrated under reduced pressure; and the residue was subjected to silica gel

column chromatography (N-heptane/ethyl acetate=10/1), thus obtaining the labeled

precursor I(III)-SPIAd-SEM-2-Larotrectinib as a white solid (20.81mg, the total yield of

21% in two steps).

[00317] Example 80

[00318] I(III)-SPIAd-SEM-5-Larotrectinib was prepared by a method similar to the

method in Example 79, and the white solid I(III)-SPIAd-SEM-5-Larotrectinib was

obtained (20mg, the total yield of 21% in two steps).

[00319] Example 81

[00320] Preparation of labeled precursor I(III)-SPIAd-THP-2-Larotrectinib

F F N0 0N

N'N O N' 0 - F 3 CCOO-I

N F3 CCOO NN N- HN N 0 0 C27 H3 0FIN 6 03 Si:620.47 C39 H 44 FIN6 0 7 Si:854.72 I -THP-2-Larotrectinib I -TfAc- THP-2-Larotrectinib

F F 00

F N O O F N' N O-0 F 3CCOO- 1 \ N H N F 3CCOO N H N kQN N

C 39H 44FIN60 7Si:854.72 H C39H 44 FIN 60 7 S1:854.72 I-TfAc- THP-2-Larotrectinib |(III)-SPIAd -THP-2-Larotrectinib

[00321] 0.13mL of dichloromethane and tert-butanol peroxide (tBuOOH, TBHP,

2.79g, 31mmol) were dosed in a 1mL reactor in a stirring way; I-THP-2-Larotrectinib

(68.25mg, 0.11mmol) was then dosed, and the resulting mixture was cooled to 0 °C in an

ice-water bath and stirred vigorously for 1 hour; TLC tracked the reaction until the raw

material was completely converted, and the obtained material was for later use.

[00322] KOH (47.12mg, 0.84mmol), dichloromethane (0.47ml), and SPIAd

becomes milky white suspension. The material obtained above for later use was dosed in

the above vigorously stirred mixture at once, the obtained mixture was continuously

stirred at 0 °C to react for 2 hours, and the reaction mixture gradually became yellow; TLC

tracked the reaction until the raw material was completely converted; the insoluble matter

was filtered off, and the reaction flask was washed three times with dichloromethane, 0.12

mL each time, the insoluble matter was rinsed with the washing liquid, and the yellow

dichloromethane liquid was collected. The collected liquid was concentrated under

reduced pressure (at 20 °C) evaporated to remove dichloromethane, and then dried for 1

hour under high vacuum, thus obtaining the labeled precursor

I(III)-SPIAd-THP-2-Larotrectinib as a yellow solid (27.08mg, 28.8%).

[00323] Example 82

[00324] I(III)-SPIAd-THP-5-Larotrectinib was prepared by a method similar to the

method in Example 81, and the white solid I(III)-SPIAd-THP-5-Larotrectinib was

obtained (27mg, 28%).

[00325] Example 83

[00326] Preparation of labeled precursor I(III)-SPIAd-EE-2-Larotrectinib

F F

N ON I__N -N F3CCOO-I F0CCO N N

0 0

C 25HaoFINeOS:608.46 C 29H 30FIN 60 7:834.49 I -EE-2-Larotrectinib I -TfAc- EE-2-Larotrectinib

F F

O 0 F 3CCOO-.._1 H F 3 CCOO NHN H 0H NKN O "' ON

C 29H3 OFIN 6 0 7:834.49 H C3 gH44FIN 6 07:842.71 I -TfAc- EE-2-Larotrectinib |(III)-SPIAd -EE-2-Larotrectinib

[00327] 0.13mL of chloroform, 0.3mL of trifluoroacetic acid, trifluoroacetic

anhydride (0.9mL), Oxone (H3K5018S4 = 614.76, 101.44mg, 0.165mmol), and

I-EE-2-Larotrectinib (71.2mg, 0.11mmol) were dosed in a 2mL reactor, the obtained

solution was cooled to 0 °C in an ice-water bath and stirred vigorously; anhydrous

potassium acetate (0.53g, 6.45mmol) was then slowly dosed in the mixture; the obtained

mixture was further stirred at 0 °C to 5 °C to react for 0.5 hour; the reaction solution was

then diluted with water and extracted with dichloromethane; the organic phases were

combined and extracted, then dried with anhydrous sodium sulfate, filtered, and

obtaining a colorless solid I-EE-TfAc-2-Larotrectinib (52.24mg, 0.064 mmol, 58%).

[00328] 10% sodium carbonate aqueous solution (w/v.0.75mL, 0.33M) containing

in a 2mL reaction flask, and then I-EE-TfAc-2-Larotrectinib (0.096mmol, 84.53mg) was

and TLC tracked the reaction until the raw material was completely converted; the

reaction mixture was diluted with 3mL of water, and extracted three times with

dichloromethane, 5ml each time; the organic extract phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure; the residue was subjected to silica gel column chromatography and eluted with n-heptane/ethyl acetate=10/1, thus obtaining the labeled precursor I(III)-SPIAd-EE-2-Larotrectinib as a white solid (40.60mg, the total yield of 43.8% in two steps).

[00329] Example 84

[00330] I(III)-SPIAd-EE-5-Larotrectinib was prepared by a method similar to the

method in Example 83, and the white solid I(III)-SPIAd-EE-5-Larotrectinib was obtained

(40mg, the total yield of 43% in two steps).

[00331] Example 85

[00332] Preparation of labeled precursor I (III)-SPIAd-Als-2-Larotrectinib

F F

/-N O O -N O O N N F3 CCOO-I O KII~hJ N HN~ 3CCOdN HN6 N. F 0 0

C 26H 30FIN 604S:656.52 C 2qH 30F 7 N 60 6S:882.55 I -Als-2-Larotrectinib I - Als-TfAc-2-Larotrectinib

F F 0 '0i

F3 CCOO-I N H O

0 ~H0 C29H 3OF 71NG0 6S:882.55 H C 37H4 0FLN608S:874.72 I -Als-TfAc-2-Larotrectinib I(III)-SPIAd -Als-2-Larotrectinib

[00333] 0.8mL of acetone was dosed in a 2 mL reactor and I-Als-2-Larotrectinib

(91.91 mg, 0.14 mmol) and mCPBA (62.86 mg, 0.22 mmol) were then dosed in a stirring

way; the mixture was stirred vigorously at 10 °C to 15 °C for 1 hour to 2 hours, 1.68mL of

SPIAd (34.5mg, 0.15mmol) dissolved in 10% sodium carbonate solution was dosed in the

reaction mixture, and the pH of the mixture was adjusted to 8 with 10% sodium carbonate

solution. The mixture was vigorously stirred at 10 °C to 15 °C for 2 hours and TLC

tracked the reaction until the raw material was completely converted; the reaction solution was transferred to a 5mL reaction flask and diluted, and then extracted three times with dichloromethane, 5 mL each time; the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure; and the residue was subjected to silica gel column chromatography

(eluted with 0-40% ethyl acetate-petroleum ether solution), thus obtaining the labeled

precursor I (III)-SPIAd-Als-2-Larotrectinib as a yellow solid (25.72mg, 21%).

[00334] Example 86

[00335] I(III)-SPIAd-Als-5-Larotrectinib was prepared by a method similar to the

method in Example 85, and the white solid I(III)-SPIAd-Als-5-Larotrectinib was obtained

(26mg, 21%).

[00336] Example 87

[00337] Preparation of labeled precursor I(III)-SPIAd-PMP-2-Larotrectinib or

F OMe F

0 __ __-Nm o\Ia o 5''NN

N N6 FCCOO N N N

0 0 C 2 8H 2 8FIN 603:642.47 C 32 H2 8F7 1N 607 :868.50 I - PMP-2-Larotrectinib I - PMP-TfAc-2-Larotrectinib

F F OMe O OMe CCOO F 3 N N HNN HN- NJ HN_ H,.

C32H 2UF71N 607:868.50 H C41H 42FIN 6 0 7 :876.72 I - PMP-TfAc-2-Larotrectinib |(Ill)-SPIAd -PMP-2-Larotrectinib

[00338] 0.53mL of trifluoroacetic acid and 0.18mL of chloroform were dosed in a 2

mL reactor; I-PMP-2-Larotrectinib (9 6. 3 7mg 0.15 mmol), sodium chloride (0.59mg,

0.Olmmol), and Oxone (H3K5018S4 = 614.76, 138.32mg, 0.225mmol) were dosed in a

tracked the reaction until the raw material was completely reacted; the reaction mixture was then concentrated under reduced pressure to remove the solvent; 1.2mL of ethanol and 0.68 mL of SPIAd (34.5mg, 0.15mmol) dissolved in 10% sodium carbonate solution were dosed in the residue, and the pH of the mixture was adjusted to 9 with the 10% sodium carbonate solution. The mixture was stirred at room temperature for 70 minutes;

TLC tracked the reaction till the end of the reaction; the reactant was diluted with 3mL of

water, and extracted three times with DCM, 5 mL each time; the organic phases were

combined, dried over anhydrous magnesium sulfate, concentrated under reduced pressure;

and the residue was subjected to silica gel column chromatography (eluted with 0-40%

ethyl acetate-petroleum ether solution), thus obtaining the labeled precursor I

(III)-SPIAd-PMP-2-Larotrectinib as a yellow solid (43.39mg, 33%).

[00339] Example 88

[00340] I(III)-SPIAd-PMP-5-Larotrectinib was prepared by a method similar to the

method in Example 87, and the white solid I(III)-SPIAd-PMP-5-Larotrectinib was

obtained (43mg, 33%).

[00341] Step 3: Preparation of radioisotope

[00342] Quaternary ammonium salt (TEABC) was used to replace K222 with

higher toxicity, the anion exchange cartridge was washed with acetonitrile to elute the

enriched [ 18F] fluoride, and dehydration drying was carried out for less than 5 minutes;

therefore, a short-time, low-toxicity, and high-radiochemical-purity preparation method

was provided. The preparation process of labeled product [ 18F]-Larotrectinib was as

follows:

[00343] Step (1). Preparation of [ 18F] fluoride target water: [ 18F] fluoride target

water was produced by 1 80(p,n) 18F nuclear reaction. GE PETTrace editcyclotron (which

can produce approximately 150 mCi [ 18F] fluoride target water with 40 A beam for 2

minutes) was used. [ 18F] Fluoride target water was delivered to a sterile lead-protected hot

cell of 180-enriched water by nitrogen pressure; [ 18F] fluoride target water produced by this method was usually further diluted with Milli-Q (Millipore ultrapure water instrument) ultra-purified water to obtain 1-3mCi/ml [ 18F] fluoride target water liquid before being used in research.

[00344] Step (2). Enrichment of [ 18F] fluoride by QMA anion exchange solid phase

extraction cartridge (QMA): An aliquot of the target water containing an appropriate

amount of [ 18F] fluoride was allowed to slowly pass through the anion exchange solid

phase extraction cartridge (QMA) under a nitrogen flow; the anion exchange solid phase

extraction cartridge (QMA) was pre-activated by being washing with NaHCO 3(aq(8.4%,

1mL) and water (20 mL, until the pH indicator showed neutrality); the [ 18F] fluoride was

enriched on the QMA anion exchange solid phase extraction cartridge (QMA), and 180

and other impurities were separated and eluted, thus obtaining [1 8F] fluorine source for the

[ 18F] fluoride QMA anion exchange solid phase extraction cartridge (QMA).

[00345] Step (3). The [ 18F] fluoride enriched on the QMA anion exchange solid

phase extraction cartridge (QMA) was eluted to obtain a quaternary ammonium salt or

inorganic salt solution of [ 18F] fluoride; the [ 18F] fluoride enriched on QMA anion

exchange solid phase extraction cartridge (QMA) was washed with a solution obtained by

dissolving an organic or inorganic base (for example, a certain amount of

tetraethylammonium bicarbonate, e.g., 8mg) in acetonitrile and water (1mL, v/v 7:3) or

acetonitrile (1mL) or methanol (1mL) or ethanol (1mL), and the [ 18F] fluoride was eluted

into a V-shaped flask sealed with a Teflon-lined diaphragm, thus obtaining an acetonitrile

aqueous solution or an acetonitrile or methanol solution of the organic or inorganic salt of

[ 18F] fluoride.

[00346] Step (4). Preparation of dry quaternary ammonium salt or inorganic salt of

[ 18F] fluoride: the V-shaped flask containing the acetonitrile aqueous solution or

acetonitrile or methanol or ethanol solution of the organic or inorganic salt of [ 18F] fluoride and sealed with a Teflon-lined diaphragm was heated to 95 °C to 110 °C, and at the same time nitrogen gas was dried by passing through the P205-DrieriteTM column and then used to blow the V-shaped flask, and then the exhaust gas was discharged through the vented flask. When no liquid was seen in the flask, the flask was taken out of the hot bath, anhydrous acetonitrile (1 mL) was then dosed in the flask, and the flask was heated again until the flask was dry. This step was repeated another three times. Then the flask was cooled at room temperature under nitrogen flow, thus obtaining dry organic or inorganic salt of 18

[ F] fluoride, such as [F]KF/K2C3/K2.2.2, [1 F]KF/K2C204/K2.2.2, 18

[ F]KF/KOTf,

[ 18F]Et4NF, [ 18F]Et4NHCO3, [ 18F]Et4NOMs, and [18 F]Et4NOTf; its radioactive [ 18F]

fluoride recovery rate varied depending on the elution process used.

[00347] Step (5). Construction of labeled product 18[ F]-Larotrectinib reaction

system: the required solvent (e.g., DMF) was dosed in the V-shaped flask containing the

dry organic or inorganic salt of [ 18F] fluoride (activity meter measured (t)activity) to

re-dissolve the dry organic or inorganic salt, and the labeled precursor was then dosed to

take reaction under a certain condition, thus obtaining a crude reaction solution of

undeprotected labeled product [1 8F]-Larotrectinib.

[00348] Step (6). Deprotection of undeprotected labeled product 18

[ F]-Larotrectinib:

a certain amount of organic base or inorganic base or organic acid or inorganic acid was or

was not dosed in the reaction solution, and the hydroxyl protecting group was removed by

heating, thus obtaining the crude reaction solution of labeled product [ 18F]-Larotrectinib.

[00349] Step (7). Separation and purification of labeled product [ 18F]-Larotrectinib:

semi-preparative HPLC or Waters Sep-Pak C-18 cartridge purification was carried out to

prepare high-purity labeled product [ 18F]-Larotrectinib, and the labeled product

[ 18F]-Larotrectinib was rinsed with a solvent into a sterile vacuum flask, blow dried with

nitrogen at 60 °C for 20 minutes, and reconstituted with brine; the obtain solution which

included 100ul of 25% Vitamin C aqueous solution and 100ul of 20% Tween 80 ethanol solution was labeled product 18

[ F]-2-Larotrectinib injection.

[00350] Step (8). Analysis and identification of labeled product 1[8 F]-Larotrectinib:

the identity and purity (radiochemical purity and chemical purity) of the product were

determined by radio-HPLC (60:40 CH 3 CN:H 20+0.IN ammonium formate, Phenomenex

Luna C18, 250 x 4.6 mm, 5 m, UV at 254 nm; CH3CN/0.1 M NH4•HCO2(aq) (v/v, 7/3),

flow rate: 1.0 mL/min) and radio-TLC (EtOAc + 0-5% EtOH). The product had the

radiochemical purity and chemical purity of greater than 90% to 99%. The radiochemical

yield was determined as the percentage of radioactivity of the labeled precursor dosed to

the DMF-diluted [ 1 8F]Et4NHCO3 and separated as the final product from the amount of

activity in the V-shaped reaction flask, without attenuation correction. The radiochemical

yield was 20 to 45.3 (without attenuation correction), the radiochemical purity was greater

than 99%, and the specific activity was 2.56 Ci/[tmol to 18 Ci/[tmol.

[00351] The following specific examples are described for detailed explaination:

[00352] Example 89. Preparation process of organic salt or inorganic salt of [ 18F]

fluoride

[00353] An aliquot of the target water containing an appropriate amount of [ 18F]

fluoride was allowed to slowly pass through the anion exchange solid phase extraction

cartridge (QMA) under a nitrogen flow; the anion exchange solid phase extraction

cartridge (QMA) was pre-activated by being washed with NaHCO 3(aq)(8.4%, 1mL) and

water (20 mL, until the pH reached neutrality); the [ 18F] fluoride was enriched on the

QMA anion exchange solid phase extraction cartridge (QMA), and 180 and other

impurities were separated and eluted, thus obtaining [ 18F] fluorine source for the [ 18F]

fluoride QMA anion exchange solid phase extraction cartridge (QMA).

[00354] The [ 18F] fluoride enriched on QMA anion exchange solid phase extraction

cartridge (QMA) was washed with a solution obtained by dissolving different organic or inorganic bases in acetonitrile and water (1mL, v/v 7:3) or acetonitrile (1mL) or methanol

(1mL) or ethanol (1mL), and the [ 18F] fluoride was eluted into a V-shaped flask sealed

with a Teflon-lined diaphragm, thus obtaining an acetonitrile aqueous solution or an

acetonitrile or methanol solution of the organic or inorganic salt of [ 18F] fluoride.

[00355] The V-shaped flask containing the acetonitrile aqueous solution or

fluoride and sealed with a Teflon-lined diaphragm was heated to 95 °C to 110 °C, and at

the same time nitrogen gas was dried by passing through the P205-DrieriteTM column and

then used to blow the V-shaped flask, and then the exhaust gas was discharged through the

vented flask. When no liquid was seen in the flask, the flask was taken out of the hot bath,

anhydrous acetonitrile (1 mL) was then dosed in the flask, and the flask was heated again

until the flask was dry. This step was repeated another three times. Then the flask was

cooled at room temperature under nitrogen flow, thus obtaining dry organic or inorganic

salt of [ 18F] fluoride, such as [18F]KF/K2C3/K2.2.2, [ 18 F]KF/K2C204/K2.2.2, 18

[ F]KF/KOTf,

[ 18F]Et4NF, [ 18F]Et4NHCO3, [ 18F]Et4NOMs, and 18

[ F]Et4NOTf; its radioactive [ 18F]

fluoride recovery rate varied depending on the elution process used. Test data were shown

in Table 1 (Influence of different eluents and organic bases or inorganic bases on the

elution efficiency of [ 18F] fluoride) and Table 2 (Influence of different loading of organic

or inorganic bases on the elution efficiency of [ 18F] fluoride, with acetonitrile as the

elution solvent).

[00356] Table 1. Influence of different eluents and organic bases (8mg) or inorganic

bases (8mg) on the elution efficiency of [ 18F] fluoride

18F recovery K2C03/ K2C204 KOTf TEAOTf( TBAOMs( Et4NHCO3( Et4NF(T

rate (%) K222 /K222 Et4NOTf) Et4NOMs) TEABC) BAF)

Methanol 8.2 10.2 11.3 34.2 34.4 34.6 32.1

(1mL)

Ethanol 4.8 12.4 12.4 35.2 35.2 33.6 30.1

(1mL)

Acetonitrile 4.0 14.6 13.2 43.6 42.3 43.5 41.7

(1mL)

Acetonitrile 44.0 43.1 43.6 41.5 40.4 40.5 40.1

-water

(1mL)

Water 36.0 35.1 35.4 40.7 39.6 38.6 36.7 (1mL)

[00357] Table 2 Influence of organic or inorganic bases in different doses on the

elution efficiency of [ 1F] fluoride, with 1mL of acetonitrile as the elution solvent).

18F recovery K2C03/ K2C204 KOTf TEAOTf( TBAOMs( Et4NHCO3( Et4NF(T

rate (%) K222 /K222 Et4NOTf) Et4NOMs) TEABC) BAF)

1 mg 1.3 2.1 1.3 14.2 14.4 16.6 12.1

4 mg 4.0 14.6 13.2 43.6 42.3 43.5 41.7

8 mg 6.8 16.6 13.5 45.1 48.3 46.5 42.7

mg 8.0 16.8 13.4 44.7 47.5 45.5 42.1

12 mg 8.5 16.1 13.1 43.1 45.9 43.6 41.3

[00358] Example 90

[00359] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Bz-2-Larotrectinib.

F F

O K'NH OCH 18 F NN OOCC 6 H5 H N N N H 0 0 1 H C41H42FIN 6 07:876.72 C 28H26F FN 60 3: 531.55 1 1(11)-SPIAd - Bz-2-Larotrectinib F -Bz-2-Larotrectinib F F

0 OOCCeH5 F N OH F N-N 18FF N8

8 C2 sH 26 F'OFN 6 0 3 : 531.55 C2 1H 2 2F' FN 60 2 : 427.45

[00360] Solution 1. According to the preparation process of labeled product

[ 1 8F]-Larotrectinib, [ 18F] fluorine source (0.5mL, activity meter measured (t)activity:

1.5mCi) QAM[ 1 8F] fluoride was taken and then the fluorine source QAM[18F] fluoride

anion exchange solid phase extraction cartridge (QMA) was eluted into a V-shaped

reaction flask with the solution of N,N,N,N-tetraethylammonium methanesulfonate

(TBAOMs, 8.0mg) dissolved in 1mL of acetonitrile; the obtained acetonitrile solution of

[ 1 8F]Et4NOMs was dehydrated to dryness by repeated azeotropic evaporation through

anhydrous acetonitrile, and the residue was diluted with anhydrous DMF (0.4mL) to

obtain a 20mg/mL 1[ 8 F]Et4NOMs solution.

[00361] 400uL of sample was separated from the above DMF solution and dosed in

a V-shaped reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib (4.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 120 °C and reacted for 20 minutes under an airtight

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Bz-2-Larotrectinib

basically disappeared; radio-TLC tracked reaction mixture sampling (1-2uL) and it was

detected that there was the labeled product [ 18F]-Bz2-Larotrectinib.

[00362] Anhydrous K2C03(0.86mg) was dosed in the reaction mixture; the mixture

was then heated to 100 °C to react for 10 minutes; the reaction solution was cooled in a 0

°C ice bath; 10% HCl aqueous solution (15 L, 1.05 g/cm 3) was dropwise dosed until the

solution was neutralized to neutral. The reactant was further diluted with HPLC buffer

(60:40 CH3CN:H20+0.iN ammonium formate, 2mL), and passed through the Waters

C-18 Sep-Pak that had been washed sequentially with ethanol (1mL) and water (5mL).

Sep-Pak was rinsed with water (2mL), the desired product was eluted with ethanol (1mL), rinsed into a sterile vacuum flask, blow dried with nitrogen at 60 °C for 20 minutes, and reconstituted with brine; the obtain solution which included 100ul of 25% Vitamin C aqueous solution and 100ul 20% Tween 80 ethanol solution was labeled product

[1 8F]-Larotrectinib injection.

[00363] The sample was analyzed by radio-TLC (silica gel plate, 100% ethyl

acetate as developing agent) to determine the radiochemical conversion (RCC), and the

identity and purity of the product were determined by radioactive HPLC (60:40

CH3CN:H20+0.1N ammonium formate, Phenomenex Luna C-18 column) and radio-TLC

(silica gel plate, 100% ethyl acetate as developing agent). The product had the

radiochemical purity of greater than 93% and the chemical purity of greater than 95%. The

co-injection method of radioactive and non-radioactive controls was carried out, and the

double detectors of radioactive detector and non-radioactive ultraviolet detector

determined the consistency of peak position to identify the labeled product. The

radiochemical yield was determined as the percentage of radioactivity of the iodonium

precursor dosed to the DMF-diluted [1 8 F]Et4NOMs solution and separated as the final

product from the amount of activity in the V-shaped reaction flask, without attenuation

correction. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 28.56%

relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is

usually 1.35mCi), and the specific activity (18.2lCi/ mol) was obtained from the final

preparation). The deprotection rate of the product was 95%.

[00364] Solution 2. The operation process here was the same as that in Solution 1

except that DMF solution was used to separate 400uL of samples respectively containing

[1 8F]KF/K2C03/K2.2.2, [1 8F]KF/K2C204/K2.2.2, [1 8F]KF/KOTf, [18F]Et4NF, [18 F]Et4NHCO3,

[ 18F]Et4NOMs, and [ 18F]Et4NOTf(their radioactivities were all 1.35mCi), and the samples

were respectively dosed in V-shaped reaction flasks with the white solid labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib (4.0mg). The mixture was heated to 120 °C and reacted for 20 minutes under an airtight condition until TLC tracked that the labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib basically disappeared; radio-TLC tracked reaction

mixture sampling (1-2uL) and it was detected that there was the labeled product

[ 18F]-Bz-2-Larotrectinib. The subsequent steps were the same as those in Solution 1, and

the obtained product had the radiochemical purity and chemical purity of greater than 99%.

The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib relative to that in the

V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi) and the

specific activity were shown in Table 3 below.

[00365] Table 3. Influence of organic or inorganic base in different doses on the

radiochemical yield and specific activity of [ 18F]-2-Larotrectinib

K2C03/ K2C204 KOTf TEAOTf( TBAOMs( Et4NHC Et4NF

K222 /K222 Et4NOTf) Et4NOMs) 03 (TBAF)

(TE ABC)

RCC(%) 0.08 8.62 0.56 25.0 28.0 26.4 22.1

Specific

activity 0.001 0.1 0.002 12.8 18.0 14.1 11.5 (Ci/pm

ol)

[00366] Solution 3. The operation process here was the same as that in Solution 1

except that the fluorine source QAM[ 18F] fluoride anion exchange solid phase extraction

cartridge (QMA) was eluted into a V-shaped reaction flask with the solution of

N,N,N,N-tetraethylammonium methanesulfonate (TBAOMs, 8mg) dissolved in 1mL of

acetonitrile; the obtained acetonitrile solution of [18F]Et4NOMs was dehydrated to dryness

by repeated azeotropic evaporation through anhydrous acetonitrile, and the residue was

diluted with different anhydrous solvents DMF (0.4mL), DMSO (0.4mL), DMA (0.4mL),

CH3CN (0.4mL), and NMP (0.4mL) to obtain 10mg/mL 1[ 8 F]Et4NOMs solution. The subsequent steps were the same as those in Solution 1, and the obtained product had the radiochemical purity and chemical purity of greater than 99%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib relative to that in the V-shaped reaction flask

(the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi) and the specific activity were

shown in Table 4 below.

[00367] Table 4. Influence of different solvents on the radiochemical yield and

specific activity of 18

[ F]-2-Larotrectinib

DMSO DMA CH3CN NMP DMF(0.4 mL) (0.4 mL) (0.4 mL) (0.4 mL) (0.4 mL)

RCC(%) 28.0 27.2 26.1 27.8 21.0

Specific

activity 18.0 16.9 15.6 17.2 10.5 (Ci/ mol)

[00368] Solution 4. The operation process was the same as that in Solution 1 except

that the fluorine source QAM[18 F] fluoride anion exchange solid phase extraction

cartridge (QMA) was respectively eluted into a V-shaped reaction flask with the solutions

of N,N,N,N-tetraethylammonium methanesulfonate in different doses (TBAOMs, 1mg,

4.0mg, 8mg, 10mg, 12mg) dissolved in 1mL of acetonitrile; the obtained acetonitrile

solution of [1 8F]Et4NOMs was dehydrated to dryness by repeated azeotropic evaporation

through anhydrous acetonitrile, and the residue was diluted with anhydrous DMF (0.4mL)

to obtain 10mg/mL 1[8 F]Et4NOMs solution. The subsequent steps were the same as those

in Solution 1, and the obtained product had the radiochemical purity and chemical purity

of greater than 99%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib relative

to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually

1.35mCi) and the specific activity were shown in Table 5 below.

[00369] Table 5. Influence of organic or inorganic base in differnt doses on the

radiochemical yield and specific activity of [ 18F]-2-Larotrectinib

TBAOMs TBAOMs(E TBAOMs(Et4 TBAOMs(Et4 TBAOMs(Et4 (Et4NOMs) t4NOMs) 8 NOMs) NOMs) NOMs) 4 mg 1 mg mg 10 mg 12 mg

RCC(%) 0.08 8.62 28.0 25.0 28.0

Specific

activity 0.001 0.1 18.0 12.8 18.0

(Ci/ mol)

[00370] Solution 5. The operation process here was the same as that in Solution 1

except that 400uL of sample was separated from the above DMF solution and dosed in a

V-shaped reaction flask containing the white solid labeled precursor

1.35 mCi). The mixture was heated to different temperatures, e.g., 100 °C, 110 °C, 120 °C,

135 °C, 145 °C, 155 °C, and 160 °C and reacted for 20 minutes respectively under an

airtight condition until TLC tracked that the labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib basically disappeared. The subsequent steps were the

same as those in Solution 1, and the obtained product had the radiochemical purity and

chemical purity of greater than 99%. The uncorrected radiochemical yield of

[ 18F]-2-Larotrectinib relative to that in the V-shaped reaction flask (the radioactivity of

[ 18F] Et4NOMs is usually 1.35mCi) and the specific activity were shown in Table 6 below.

[00371] Table 6. Influence of different reaction temperatures on the radiochemical

yield and specific activity of [ 18F]-2-Larotrectinib

100°C 110°C 120°C 135°C 145°C 155°C 160°C

RCC(%) 24.2 26.1 28.0 28.0 28.0 28.0 28.0

Specific

activity 14.3 15.6 18.0 18.0 18.0 18.0 18.0

(Ci/ mol)

[00372] Solution 6. The operation process here was the same as that in Solution 1 except that 400uL of sample was separated from the above DMF solution and dosed in

V-shaped reaction flasks containing the white solid labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib in different doses (0.5mg, 1.0mg, 2.0mg, 4.0mg, 6mg)

(the radioactivity of [ 18F]Et4NOMs is usually 1.35 mCi). The mixture was heated to 145

°C and reacted for 20 minutes under an airtight condition until TLC tracked that the

labeled precursor I(III)-SPIAd-Bz-2-Larotrectinib basically disappeared. The subsequent

steps were the same as those in Solution 1, and the obtained product had the radiochemical

purity and chemical purity of greater than 99%. The uncorrected radiochemical yield of

[ 18F] Et4NOMs is usually 1.35mCi) and the specific activity were shown in Table 7 below.

[00373] Table 7. Influence of labeled precursor in different doses on the

radiochemical yield and specific activity of [ 18F]-2-Larotrectinib

Labeled Labeled Labeled Labeled Labeled

precursor precursor precursor precursor precursor

0.5mg 1.0 mg 2.0 mg 4.0 mg 6.0 mg

RCC(%) 26.8 28.1 28.2 28.2 28.3

Specific

activity 16.1 18.0 18.1 17.6 17.4

(Ci/ mol)

[00374] Solution 7. The operation process here was the same as that in Solution 1

V-shaped reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib (0.5 mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was respectively heated to 120 °C, 135 °C, 145 °C, 155 °C, and

160 °C and reacted for different times, e.g., 5 minutes, 10 minutes, 15 minutes, and 20

minutes under an airtight condition until TLC tracked that the labeled precursor

chemical purity of greater than 99%. The uncorrected radiochemical yield of

[ 1 8F]-2-Larotrectinib relative to that in the V-shaped reaction flask (the radioactivity of

[ 18F] Et4NOMs is usually 1.35mCi) and the specific activity were shown in Table 8 below.

[00375] Table 8. Influence of different labeling reaction times on the radiochemical

yield and specific activity of 1[ 8 F]-2-Larotrectinib

Temperature Specific activity Time (min) RCC(%) (C) (Ci/ mol)

12 15.3 7.6

15 17.9 10.2 120 °C 18 23.4 14.2

20 28.0 18.0

12 18.4 9.7

15 23.8 14.5 135 °C 18 28.0 18.0

20 28.0 18.0

8 17.6 9.3

12 23.7 14.6

145 °C 15 28.0 18.0

18 28.0 18.0

20 27.8 15.6

8 18.3 9.8

10 24.6 13.8

155 °C 12 28.0 18.0

15 28.0 17.1

18 27.5 16.6

160 °C 5 13.5 6.7

8 16.8 8.9

12 27.6 15.4

15 27.7 14.9

18 26.3 14.3

[00376] Solution 8. According to the preparation process of labeled product

[ 18F]-Larotrectinib, [ 18F] fluorine source (0.5mL, activity meter measured (t)activity:

1.5mCi) QAM[ 18F] fluoride was taken and then the fluorine source QAM[18F] fluoride

reaction flask with the solution of N,N,N,N-tetraethylammonium methanesulfonate

[ 18F]Et4NOMs was dehydrated to dryness by repeated azeotropic evaporation through

obtain a 20mg/mL 18

[ F]Et4NOMs solution.

[00377] 400uL of sample was separated from the above DMF solution and dosed in

a V-shaped reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 155 °C and reacted for 12 minutes under an airtight

detected that there was the labeled product [ 18F]-Bz-2-Larotrectinib.

[00378] Anhydrous potassium tert-butoxide (KOtBu, 0.70mg) was dosed in the

reaction mixture; the mixture was then heated to 80 °C to react for 8 minutes; the reaction

solution was cooled in a 0 °C ice bath; 10% HCl aqueous solution (15 L, 1.05 g/cm3) was

dropwise dosed until the solution was neutralized to neutral. The reactant was further

diluted with HPLC buffer (60:40 CH3CN:H20+0.iN ammonium formate, 2mL), and

passed through the Waters C-18 Sep-Pak that had been washed sequentially with ethanol

(1mL) and water (5mL). Sep-Pak was rinsed with water (2mL), the desired product was

eluted with ethanol (1mL), rinsed into a sterile vacuum flask, blow dried with nitrogen at

°C for 20 minutes, and reconstituted with brine; the obtain solution which included

100ul of 25% Vitamin C aqueous solution and 100ul 20% Tween 80 ethanol solution was

labeled product [1 8 F]-2-Larotrectinib injection.

[00379] The sample was analyzed by radio-TLC (silica gel plate, 100% ethyl

identity and purity of the product were determined by radioactive HPLC (60:40

CH3CN:H20+0.1N ammonium formate, Phenomenex Luna C-18 column) and radio-TLC

(silica gel plate, 100% ethyl acetate as developing agent). The product had the

deprotection rate of 100% and the radiochemical purity and chemical purity of greater

than 99%. The co-injection method of radioactive and non-radioactive controls was

carried out, and the double detectors of radioactive detector and non-radioactive ultraviolet

detector determined the consistency of peak position to identify the labeled product. The

correction. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 41.82%

usually 1.35mCi), and the specific activity (27.11Ci/ mol) was obtained from the final

preparation).

[00380] Example 91

[00381] Similar to the method of Example 90, the labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Bz-5-Larotrectinib, the product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected radiochemical yield of [1 8F]-5-Larotrectinib was 41.15% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity (27.13Ci/ mol) was obtained from the final preparation).

[00382] Example 92

[00383] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-TBS-2-Larotrectinib.

HN NN OTBS - N OTBS

18 H C 40H 50FN,06Si:884.86 C27 Ha6 F FN 602Si:541.71 |(Ill)-SPIAd -TBS -2-Larotrectinib isF -TBS-2- Larotrectinib ~JN~IN H 'OO

F 0" F 0

NN OTBS NN OH 18F N s

8 8 C 27 HasF' FN6 2 Si:541.71 0 C2 1H2F' FN 6 02 : 427.45 1 sF -TBS-2- Larotrectinib IOF - 2-Larotrectinib

[00384] According to the preparation process oflabeled product [ 18 F]-Larotrectinib,

[ 1 8 F]fluorine source(0.5mL,activitymetermeasured (t)activity: 1.5mCi)QAM[F]

fluoride was taken and then the fluorine source QAM[ 8 F] fluoride anion exchange solid

phase extraction cartridge (QMA) was eluted into aV-shaped reaction flask with the

solution of N,N,N,N-tetraethylammonium methanesulfonate (TBAOMs, 8.0mg) dissolved

in1mL of acetonitrile; the obtained acetonitrile solution of [ 18 F]Et4NOMs was dehydrated

todrynessby repeated azeotropic evaporation through anhydrousacetonitrile,andthe

residue was diluted withanhydrous DMF (0.4mL) tofobtainda 20mg/mni[F]Et4NOMs

solution.

[00385] 400uL of sample was separated from the above DMF solution and dosed in a V-shaped reaction flask containing the white solid labeled precursor

I(III)-SPIAd-TBS-2-Larotrectinib (1.0mg) (the radioactivity of [1 8F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-TBS-2-Larotrectinib

detected that there was the labeled product 1[ 8F]-2-Larotrectinib.

[00386] The reactant was further diluted with HPLC buffer (60:40

CH3CN:H20+0.1N ammonium formate, 2mL), and passed through the Waters C-18

Sep-Pak that had been washed sequentially with ethanol (1mL) and water (5mL). Sep-Pak

was rinsed with water (2mL), the desired product was eluted with ethanol (1mL), rinsed

into a sterile vacuum flask, blow dried with nitrogen at 60 °C for 20 minutes, and

reconstituted with brine; the obtain solution which included 100ul of 25% Vitamin C

aqueous solution and 100ul of 20% Tween 80 ethanol solution was labeled product

[1 8F]-2-Larotrectinib injection.

[00387] The sample was analyzed by radio-TLC (silica gel plate, 100% ethyl

identity and purity of the product were determined by radio-HPLC (60:40

CH3CN:H20+0.1N ammonium formate, Phenomenex Luna C-18 column) and radio-TLC

(silica gel plate, 100% ethyl acetate as developing agent). The product had the

radiochemical purity and chemical purity of greater than 99%. The co-injection method of

radioactive and non-radioactive controls was carried out, and the double detectors of

radioactive detector and non-radioactive ultraviolet detector determined the consistency of

peak position to identify the labeled product. The radiochemical yield was determined as

the percentage of radioactivity of the iodonium precursor dosed to the DMF-diluted

[18F]Et4NOMssolution and separated as the final product from the amount of activity in

the V-shaped reaction flask, without attenuation correction. The uncorrected radiochemical yield of [ 8 F]-2-Larotrectinib was 45.82% relative to that in the V-shaped reaction flask

(the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity

(29.82Ci/ mol) was obtained from the final preparation).

[00388] Example 93

[00389] Similar to the method of Example 92, labeled product 1[8 F]-5-Larotrectinib

was prepared from the labeled precursor I(III)-SPIAd-TBS-5-Larotrectinib. The product

had the deprotection rate of 100% and the radiochemical purity and chemical purity of

greater than 99%. The uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was

45.21% relative to that in the V-shaped reaction flask (the radioactivity of 1[ 8 F] Et4NOMs

is usually 1.35mCi), and the specific activity (29.81Ci/ mol) was obtained from the final

preparation).

[00390] Example 94

[00391] Labeled product [ 18F]2-Larotrectinib was prepared from the labeled

precursor I(III)-SPIAd-SEM-2-Larotrectinib.

F F

0 N'N OSEM N OSEM

H H OCN N S 18F Nj N

H 0 0 18 H 7 Si:900.86 C 4 0 HSOFIN 6 0 C 2 7H3 6 F FN 60 3 Si:557.71 I(Ill)-SPIAd - SEM-2-Larotrectinib 1 F -SEM- 2-Larotrectinib F F

N.N OSEM N-N OH 18 F 18F N N , 1HN Nr 0-i 1 1 C 27H 36F FN,0 3 Si:557.71 C21 H 22F FN 6 0 2 : 427.450 18 18 F -SEM- 2-Larotrectinib F -2- Larotrectinib

[00392] The operation in this example was the same as that in Example 91. The

product has the deprotection rate of 100% or above and the radiochemical purity and

chemical purity of greater than 99%. The uncorrected radiochemical yield of

[ 1 8F]-2-Larotrectinib was 45.82% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity

(29.82Ci/ mol) was obtained from the final preparation).

[00393] Example 95

[00394] Similar to the method of Example 94, labeled product 1[8 F]-5-Larotrectinib

was prepared from the labeled precursor I(III)-SPIAd-SEM-5-Larotrectinib. The product

45.55% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs

is usually 1.35mCi), and the specific activity (29.80Ci/ mol) was obtained from the final

preparation).

[00395] The product was re-sampled and detected by radio-TLC until deprotection

was completed. The reaction mixture was neutralized with 5N sodium hydroxide (5N,

0.2mL) and diluted with HPLC mobile phase (10% ethanol, 28mM hydrochloric acid,

nM ammonium acetate, pH2, 0.5mL). The diluted solution was injected into a

semi-preparative HPLC (static phase: Hamilton PRP-1, 250x 10 mm, 10 um; mobile phase

same as above, 3.5mL/min, tR = -17min), the eluent containing the product was collected

step by step; the activity was detected by an activity meter, the separation rate of 80%

(without attenuation correction) was also calculated, and the total time (from to to the end

of the synthesis EOS) 75 2 minutes. The product has the radiochemical purity and

chemical purity of 95% to 99% and the deprotection conversion rate of 98% or above.

[00396] The mixture (2uL) was subjected to TLC thin-plate chromatography

(developing agent: 100% ethyl acetate) to determine the labeling rate (RCC), then diluted

with 15 mL of water, passed through a solid phase extraction cartridge with C18 filler,

eluted with water (24 mL), and rinsed with acetonitrile (1.5 mL), and the resulting mixture

was collected in a micropycnometer.

[00397] The mixture (20uL) and fluorine-19 standard control ( 19F-Larocetrinib) were co-injected into radio-HPLC to determine the18 F-labeled product 1( 8F-Larocetrinib).

The residual reaction mixture was neutralized with 5N sodium hydroxide (5N, 0.2mL) and

diluted with HPLC mobile phase (10% ethanol, 28mM hydrochloric acid, 20nM

ammonium acetate, pH2, 0.5mL). The diluted solution was injected into a

[00398] Example 96

[00399] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-TBDPS-2-Larotrectinib.

F F O N'N OTBDPS /-N OTBDPS

H H NN19F NN

H 0 0 9 H C 50H 4FIN 60 6S: 1009.01 C 37H 40F FIN 60 2SI:666.85 1(11)-SPAd -TBDPS -2- Larotrectinib 19 F -TBDPS-2-Larotrectinib

F F

N OTBDPS N OH 19 F N N N F

9 C3 7H 4 0FFIN 60 2Si:666.85 C 21H 22F FIN102 :428.44 19 19F -TBDPS-2-Larotrectinib F -2- Larotrectinib

[00400] The operation in this example was the same as that in Example 92. The

deprotection conversion rate was 100% or above. The product had the radiochemical

[ 1 8F]-2-Larotrectinib was 45.80% relative to that in the V-shaped reaction flask (the

radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity

(29.84Ci/ mol) was obtained from the final preparation).

[00401] Example 97

[00402] Similar to the method of Example 96, labeled product 1[8 F]-5-Larotrectinib

was prepared from the labeled precursor I(III)-SPIAd-TBDPS-5-Larotrectinib. The

product had the deprotection rate of 100% and the radiochemical purity and chemical

purity of greater than 99%. The uncorrected radiochemical yield of [ 18F]-5-Larotrectinib

was 45.50% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F]

Et4NOMs is usually 1.35mCi), and the specific activity (29.83Ci/ mol) was obtained

from the final preparation).

[00403] Example 98

[00404] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-TIPS-2-Larotrectinib.

F F

O f - N OTIPS 19 -N OTIPS H S F F N Hm ONN N / N N F iHN (\H N6

19 H C43H 56 FIN 6O6SI: 926.94 CyH 42 F FIN10 2Si:584.79 |(Ill)-SPIAd -TIPS - 2-Larotrectinib 1 9F -TIPS-2-Larotrectinib

F F N'N OTIPS N OH

19F N1N9F NN

0 0 C3 7H 42FISFIN 602Si:584.79 C 21 H22FIFIN 60 2:428.44 19 F -TIPS-2-Larotrectinib 19F - 2-Larotrectinib

[00405] The operation in this example was the same as that in Example 92. the

deprotection conversion rate of 100% or above. The product had the radiochemical purity and chemical purity of greater than 99%. The uncorrected radiochemical yield of

[' 8F]-2-Larotrectinib was 45.89% relative to that in the V-shaped reaction flask (the

radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity

(29.82Cimol) was obtained from the final preparation).

[00406] Example 99

[00407] Similar to the method of Example 98, labeled product 1[8 F]-5-Larotrectinib

was prepared from the labeled precursor I(III)-SPIAd-TIPS-5-Larotrectinib. The product

45.23% relative to that in the V-shaped reaction flask (the radioactivity of 1[ 8 F] Et4NOMs

is usually 1.35mCi), and the specific activity (29.31Ci/ mol) was obtained from the final

preparation).

[00408] Example 100

[00409] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Teoc-2-Larotrectinib.

F F 0 0

0O,/ N'N OA O N.N OA

HON SF N

'H O O H C 40 H4 SFIN 60s Si:914.85 Cz7 H34F'FN60 4 Si:571.69 18 |(III)-SPIAd -Teoc-2-Larotrectinib F -Teoc- 2-Larotrectinib

F F 0

CjNNN 93 N 6

[00410] The operation in this example was the same as that in Example 92. The

product had the deprotection conversion rate of 100% or above and the radiochemical

[ 1 8F]-2-Larotrectinib was 45.82% relative to that in the V-shaped reaction flask (the

radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity

(29.8OCi/mol) was obtained from the final preparation).

[00411] Example 101

[00412] Similar to the method of Example 100, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-TPIS-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.27% relative to that in the V-shaped

reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific

activity (29.31Ci/umol) was obtained from the final preparation).

[00413] Example 102

[00414] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursor I(III)-SPIAd-Piv-2-Larotrectinib.

F F

0 0 X_ NAJ 00

H O N 6 N N HN6N N HN-, 18 H C39H"FIN6O 7: 854.72 C26H 30F FIN 603:511.56 I(III)-SPIAd -Piv-2- Larotrectinib I - PIv-2-Larotrectinib

F F 0

18iF N__N 18F QN N NH 0 OH

18 18 C 2 6H 30 F FIN 6 03:511.56 C21 H22F FIN 602:427.45 I -Piv-2-Larotrectinib 'OF -2- Larotrectinib

[00415] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

400uL of sample was separated from the above DMF solution and dosed in a V-shaped

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Piv-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Piv-2-Larotrectinib

detected that there was the labeled product [ 18F]-Piv-2-Larotrectinib.

[00416] Sodium methoxide powder (NaOMe, 0.71mg) was dosed in the reaction

mixture; the mixture was then heated to 90 °C to react for 8 minutes; the reaction solution

was cooled in a 0 °C ice bath; 10% HC aqueous solution was dropwise dosed until the

solution was neutralized to neutral. The subsequent operation in this example was the

same as that in Example 92. The product had the deprotection conversion rate of 100% or

above and the radiochemical purity and chemical purity of greater than 99%. The

uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 45.83% relative to that in the

V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the

specific activity (29.85Ci/ mol) was obtained from the final preparation).

[00417] Example 103

[00418] Similar to the method of Example 102, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Piv-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.64% relative to that in the V-shaped

activity (29.44Ci/ mol) was obtained from the final preparation).

[00419] Example 104

[00420] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Ac-2-Larotrectinib.

F F

O N' O N-N O C H 18F#C / - 0C

H, 0 N C31F N N6 H

H C36H 3 FIN 6 07: 812.64 1 C 2aH24 F FIN 603 :469.48 |(Ill)-SPIAd -Ac - 2-Larotrectinib 18F -Ac-2-Larotrectinib

F F

N N OH 18 18 F N N H3 F N

HN__HN

0 O 18 C 23H 24F FIN 6 03:469.48 C 23H 24F"FIN 80 2:427.45 1 18 F - Ac-2-Larotrectinib F- 2-Larotrectinib

[00421] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor I(III)-SPIAd-Ac-2-Larotrectinib

(1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually 1.35 mCi). The mixture was heated

to 155 °C and reacted for 12 minutes under an airtight condition until TLC tracked that the

labeled precursor I(III)-SPIAd-Ac-2-Larotrectinib basically disappeared; radio-TLC

tracked reaction mixture sampling (1-2uL) and it was detected that there was the labeled product [1 8F]-Ac-2-Larotrectinib.

[00422] Sodium methoxide powder (NaOMe, 0.11mg) was dosed in the reaction

mixture; the mixture was then heated to 80 °C to react for 8 minutes; the reaction solution

above and the radiochemical purity and chemical purity of greater than 99%. The

uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 45.84% relative to that in the

specific activity (29.88Ci/ mol) was obtained from the final preparation).

[00423] Example 105

[00424] Similar to the method of Example 104, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Ac-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.54% relative to that in the V-shaped

activity (29.25Ci/ mol) was obtained from the final preparation).

[00425] Example 106

[00426] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Tf-2-Larotrectinib.

F F

0O_ N.-N OA 18 o N'N OA H N CF 3 F CF 3

H F

H C36H 35F4 |N 60 7: 866.61 18 C23H 21F 4 FINS0 3:523.45 I(Ill)-SPIAd -Tf -2-Larotrectinib 18 F-Tf-2-Larotrectinib

F F

N O C -N OH 1 18 F N

HN-_ ( HN-, 0 0 18 1 C 23H 21F 4 FIN 803:523.45 C23 H24 F FIN 602:427.45 18 18F - Tf-2-Larotrectinib F- 2-Larotrectinib

[00427] According to the preparation process of labeled product [18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor I(III)-SPIAd-Tf-2-Larotrectinib

labeled precursor I(III)-SPIAd-Tf-Larotrectinib basically disappeared; radio-TLC tracked

reaction mixture sampling (1-2uL) and it was detected that there was the labeled product

[1 8F]-Tf-2-Larotrectinib.

[00428] Sodium ethoxide powder (NaOEt, 0.15mg) was dosed in the reaction

mixture; the mixture was then heated to 80 °C to react for 6 minutes; the reaction solution

above and the radiochemical purity and chemical purity of greater than 99%. The

uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 45.45% relative to that in the

specific activity (29.65Ci/ mol) was obtained from the final preparation).

[00429] Example 107

[00430] Similar to the method of Example 106, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Tf-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.78% relative to that in the V-shaped

activity (29.54Ci/ mol) was obtained from the final preparation).

[00431] Example 108

[00432] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-CAc-2-Larotrectinib.

F F 0 0

1 / N CHCI -N N CH

H~ O 0(CT HNL}

H C 36H37CIFIN60 7:847.08 8 C 23H23CIF FIN60 3:503.93 1I(1 -SPIAd-CAc -2-Larotrectinib 18 F-CAc-2-Larotrectini b

F F

/ OA N*/ OH 18 18 F CH 2 CI FN

HN__W, HN_

18 C 23 H2 3CIF FI10 3:503.93 C2 3H 24F'OF1N 60 2:427.45 18 F -CAc-2-Larotrectinib ISF-2- Larotrectinib

[00433] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-CAc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 155 °C and reacted for 12 minutes under an airtight condition until TLC tracked that the labeled precursor I(III)-SPIAd-CAc-2-Larotrectinib basically disappeared; radio-TLC tracked reaction mixture sampling (1-2uL) and it was detected that there was the labeled product 1[8 F]-CAc-2-Larotrectinib.

[00434] Sodium ethoxide powder (NaOEt, 0.15mg) was dosed in the reaction

above and the radiochemical purity and chemical purity of greater than 99%. The

specific activity (29.87Ci/ mol) was obtained from the final preparation).

[00435] Example 109

[00436] Similar to the method of Example 108, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-CAc-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.25% relative to that in the V-shaped

activity (29.12Ci/ mol) was obtained from the final preparation).

[00437] Example 110

[00438] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-DCAc-2-Larotrectinib.

F F 0 0

H NO 2'CHCl 2

1 NHC N H, 0 N HN H0 100 H 18 H C36H 36Cl2FIN 607:881.52 C23H 22C1 2F FIN60 3 :53$.37 IIIlI-mRPlAd -'lrA-9I nrntinih C2H2C 2 F.F I :583

F F

18F N N OCHCl 1 N OH

C C ~HN_ 18 1 C23 H22Cl 2F FIN 60 3:538.37 C 23H 24F aFINr02 :427.45 18 F-DCAc-2-Larotrectinib 1 sF-2-Larotrectinib

[00439] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-DCAc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-DCAc-2-Larotrectinib

detected that there was the labeled product [ 18F]-DCAc-2-Larotrectinib.

[00440] Sodium methoxide powder (NaOMe, 0.11mg) was dosed in the reaction

mixture; the mixture was then heated to 80 °C to react for 5 minutes; the reaction solution

above and the radiochemical purity and chemical purity of greater than 99%. The

uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 45.85% relative to that in the

specific activity (29.86Ci/ mol) was obtained from the final preparation).

[00441] Example 111

[00442] Similar to the method of Example 110, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-DCAc-5-Larotrectinib. The product had the deprotection rate of 100% and

the radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.53% relative to that in the V-shaped

activity (29.24Ci/ mol) was obtained from the final preparation).

[00443] Example 112

[00444] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Moc-2-Larotrectinib.

F F 0 \/ - OA O0 O C N O 18F N N'N Hm, H NN

H 0 H C3aH 38FINO : 828.64 1 C 23H24F sFIN 604 :485.48 |(III)-SPAd -Mc-2- Larotrectinib 'OF - Moc-2-Larotrectinib

F F 0

-N OA NN OH

18F N18OiF NN O N OH N

0 0 18 18 C 23H24F FIN 6O4:485.48 C 23H 24F FIN 602 :427.45 18 1sF - Moc-2-Larotrectinib F- 2-Larotrectinib

[00445] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Moc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 155 °C and reacted for 12 minutes under an airtight condition until TLC tracked that the labeled precursor I(III)-SPIAd-Moc-2-Larotrectinib basically disappeared; radio-TLC tracked reaction mixture sampling (1-2uL) and it was detected that there was the labeled product 1[8 F]-Moc-2-Larotrectinib.

[00446] Potassium hydroxide (KOH, 0.37mg) was dosed in the reaction mixture;

the mixture was then heated to 100 °C to react for 12 minutes; the reaction solution was

cooled in a 0 °C ice bath; 10% HCl aqueous solution was dropwise dosed until the

same as that in Example 92. The product had the deprotection conversion rate of 89.8% or

above, the radiochemical purity of 96.3% and the chemical purity of greater than 91.3%.

The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 35.45% relative to that

in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi),

and the specific activity (22.33Ci/ mol) was obtained from the final preparation).

[00447] Example 113

[00448] Similar to the method of Example 112, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Moc-5-Larotrectinib. The product had the deprotection rate of 89% and the

radiochemical purity of greater than 96% and chemical purity of greater than 91%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 35.56% relative to that in the

specific activity (22.23Ci/ mol) was obtained from the final preparation).

[00449] Example 114

[00450] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Eoc-2-Larotrectinib.

F F

O N-'N OA 0 N'N O-/ 0

10 18F ~J

F F

' ~ -N O~ 0 1 N'N OH 1 18 F 1 JN 3 N J 1F N N

0 0 18 18 C 24H 26F FIN 6O 4 :499.51 C 23H24F FIN6O 2:427.45 1 18 SF-Eoc-2-Larotrectinib F-2-Larotrectinib

[00451] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Eoc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Eoc-2-Larotrectinib

detected that there was the labeled product [ 18F]-Eoc-2-Larotrectinib.

[00452] Potassium hydroxide (KOH,0.37mg) was dosed in the reaction mixture; the

mixture was then heated to 100 °C to react for 12 minutes; the reaction solution was

same as that in Example 92. The product had the deprotection conversion rate of 90.2% or

above, the radiochemical purity of 97.5% and the chemical purity of greater than 90.1%.

The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 32.15% relative to that

and the specific activity (21.43Ci/ mol) was obtained from the final preparation).

[00453] Example 115

[00454] Similar to the method of Example 114, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Eoc-5-Larotrectinib. The product had the deprotection rate of 90% and the

radiochemical purity of greater than 97% and chemical purity of greater than 90%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 32.35% relative to that in the

specific activity (21.14Ci/ mol) was obtained from the final preparation).

[00455] Example 116

[00456] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Boc-2-Larotrectinib.

F F 0~ 0

O N'N O ON OA

N 18 F NN N HNN0 N HN6

H C39H4FINOOS:870.72 C26H 30F'OFIN 60 4:527.56 I(III)-SPIAd -Boc-2- Larotrectinib 1OF - Boc-2-Larotrectinib

F F 0 -N 0 O -N OH 18F NO F N8F

O 0 18 18 C 26H 30 F FIN 604 :527.56 C23 H24F FIN 602:427.45 18 1 F-Boc-2-Larotrectinib F-2-Larotrectinib

[00457] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Boc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Boc-2-Larotrectinib

detected that there was the labeled product 1[8 F]-Boc-2-Larotrectinib.

[00458] Trifluoroacetic acid (CF3COOH, 0.75mg) was dosed in the reaction

mixture; the mixture was then heated to 100 °C to react for 6 minutes; the reaction

solution was cooled in a 0 °C ice bath; 10% HCl aqueous solution was dropwise dosed

until the solution was neutralized to neutral. The subsequent operation in this example was

the same as that in Example 92. The product had the deprotection conversion rate of 100%

or above, the radiochemical purity of 99% and the chemical purity of greater than 99%.

The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 42.35% relative to that

and the specific activity (28.25Ci/ mol) was obtained from the final preparation).

[00459] Example 117

[00460] Similar to the method of Example 116, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Boc-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity and chemical purity of greater than 99%. The uncorrected

radiochemical yield of [ 18F]-5-Larotrectinib was 45.35% relative to that in the V-shaped

activity (29.51Ci/ mol) was obtained from the final preparation).

[00461] Example 118

[00462] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Troc-2-Larotrectinib.

F F

O N'N O HN O CC13N'O 18 OC /H- F

,Ai(J N N6J106 (J N6 (CC3

F F 0 -N O- N OH 18 NNC HI N6§CCI3 s b N N-\ (j 18F NN

0 0 18 18 C 24H 23CiF FIN 604 :602.84 C 2,H 24 F FIN102 :427.45 18 18 F- Troc-2-Larotrectinib F- 2-Larotrectinib

[00463] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Troc-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Troc-Larotrectinib

detected that there was the labeled product [ 18F]-Troc-2-Larotrectinib.

[00464] Acetic acid (CH3COOH, 0.35mg) and zinc powder (Zn, 0.82mg) were

dosed in the reaction mixture; the mixture was then heated to 100 °C to react for 8 minutes;

the reaction solution was cooled in a 0 °C ice bath; 10% HCl aqueous solution was

dropwise dosed until the solution was neutralized to neutral. The subsequent operation in

this example was the same as that in Example 92. The product had the deprotection

conversion rate of 95.8% or above, the radiochemical purity of 97.6% and the chemical

purity of greater than 95%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib

was 38.63% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F]

Et4NOMs is usually 1.35mCi), and the specific activity (25.43Ci/ mol) was obtained from the final preparation).

[00465] Example 119

[00466] Similar to the method of Example 118, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Troc-5-Larotrectinib. The product had the deprotection rate of 95% and the

radiochemical purity of greater than 97.5% and chemical purity of greater than 97.5%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 38.35% relative to that in the

specific activity (25.41Ci/ mol) was obtained from the final preparation).

[00467] Example 120

[00468] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-PMB-2-Larotrectinib.

F F

OOMe NOMe N'N 1 N F0_N N 0 18 HH F

Ha N HN f H C 4 2 H4IFINO 7 :89.75 C29 H3 0 F 0 3 :547.60 6H2F1N I(I )-SPIAd-PMB-2-Larotrectinib 1(111-SPIAdF-MB-2-Larotrctinib

F F t\ OMe - -N 0 0\_/ N -N OH 18 F F

0 0 18 C 29 H3 0 F'BFIN6 0 3 :547.60 C2,HsF FIN10 2:427.45 I(111-SPIAd -PMB-2-Larotrectinib IOF- 2-Larotrectinib

[00469] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [ 1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-PMB-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-PMB-2-Larotrectinib

detected that there was the labeled product 1[8 F]-PMB-2-Larotrectinib.

[00470] 2,3-Dichloro-5,6-dicyano-p-benzoquinone (DQQ, 0.0033mmol, 0.75mg)

was dosed in the reaction mixture; the mixture was then heated to 100 °C to react for 12

minutes; the reaction solution was cooled in a 0 °C ice bath; 10% NaHCO3 aqueous

solution was dropwise dosed until the solution was neutralized to neutral. The subsequent

operation in this example was the same as that in Example 92. The product had the

deprotection conversion rate of 91.7% or above, the radiochemical purity of 95.6% and

the chemical purity of greater than 90.7%. The uncorrected radiochemical yield of

[18F]-2-Larotrectinib was 37.43% relative to that in the V-shaped reaction flask (the

radioactivity of [ 18F] Et4NOMs is usually 1.35mCi), and the specific activity (23.12

Ci/ mol) was obtained from the final preparation).

[00471] Example 121

[00472] Similar to the method of Example 120, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-PMB-5-Larotrectinib. The product had the deprotection rate of 95% and the

specific activity (25.4OCi/ mol) was obtained from the final preparation).

[00473] Example 122

[00474] Labeled product [ 1 8F]-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-PMP-Larotrectinib.

F F

O-' OMe N'- OMe

18 N;N- F N Nom

0 OMe

O

8 1 C 28 H2 8 F' FIN 6 O3 :533.57 C2 8H24F FIN6 O2:427.45 1 I(III)-SPIAd-PMP-2-Larotrectinib SF-2-Larotrectinib

[00475] According to the preparation process oflabeled product [ 18 F]-Larotrectinib,

anhydrous DMF-dissolved [1 8 F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

400uL of sample was separated from the above DMF solution and dosed ina V-shaped

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-PMP-2-Larotrectinib (1.0mg) (the radioactivity of [ 18 F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 155°C and reacted for 12 minutes under an airtight

condition until TLC tracked that the labeled precursor I(III)-SPIAd-PMP-2-Larotrectinib

detected that there was the labeled product [ 18 F]-PMP-2-Larotrectinib.

[00476] Cerium ammonium nitrate (CAN, 0.0033mmol, 1.81mg) was dosed in the

reaction mixture; the mixture was then heated to 100°C to react for 12 minutes; the

reaction solution was cooled in a00 Cicebath;100NaHCO3 aqueous solution was

this example was the same as that inExample 92.The product had thedeprotection

conversion rate of 92.2100or above, the radiochemical purity of 95.8% and the chemical

purity of greater than 91.1%o.The uncorrected radiochemical yield of [ 8 F]-2-Larotrectinib

was 37.8700 relative to that inthe V-shaped reaction flask (theradioactivity of[ 18 F]

Et4NOMs is usually 1.35mCi), and the specific activity (23.52Ci/ mol) was obtained from

the final preparation).

[00477] Example 123

[00478] Similar to the method of Example 122, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-PMP-5-Larotrectinib. The product had the deprotection rate of 93% and the

radiochemical purity of greater than 95.5% and chemical purity of greater than 91.5%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 37.65% relative to that in the

specific activity (23.81Ci/ mol) was obtained from the final preparation).

[00479] Example 124

[00480] Labeled product [ 1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-MOM-2-Larotrectinib.

F F O N' N OMOM N OMOM H / OMOMN

HN oH N N6 -1 F N N6

1 H C 36 H 40FIN6 0 7 :814.65 C 23H 2 F FIN 603:471.50 I(Ill)-SPIAd - MOM-2-Larotrectinib 18 F-MOM- 2-Larotrectinib

F F

18 N OMOM -N OH F 'OF K I N K 1 N HN6 6 N 0 0 18 18 C2,H 2 6F FIN10 3:471.50 C 23H 24F FIN10 2:427.45 18 1 F-MOM-2-Larotrectinib F-2-Larotrectinib

[00481] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-MOM-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

1.35 mCi). The mixture was heated to 155 °C and reacted for 12 minutes under an airtight condition until TLC tracked that the labeled precursor I(III)-SPIAd-MOM-2-Larotrectinib basically disappeared; radio-TLC tracked reaction mixture sampling (1-2uL) and it was detected that there was the labeled product 1[8 F]-MOM-2-Larotrectinib.

[00482] 6N hydrochloric acid (0.0033mmol, 0.65mg) was dosed in the reaction

mixture; the mixture was then heated to 80 °C to react for 12 minutes; the reaction

solution was cooled in a 0 °C ice bath; 10% NaHCO3 aqueous solution was dropwise

dosed until the solution was neutralized to neutral. The subsequent operation in this

example was the same as that in Example 92. The product had the deprotection conversion

rate of 92.4% or above, the radiochemical purity of 95.2% and the chemical purity of

greater than 91.4%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was

36.35% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs

is usually 1.35mCi), and the specific activity (22.12Ci/ mol) was obtained from the final

preparation).

[00483] Example 125

[00484] Similar to the method of Example 124, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-MOM-5-Larotrectinib. The product had the deprotection rate of 93% and the

radiochemical purity of greater than 95.8% and chemical purity of greater than 91.8%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 36.25% relative to that in the

specific activity (22.27Ci/ mol) was obtained from the final preparation).

[00485] Example 126

[00486] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-MEM-2-Larotrectinib.

F F O N'N OMEM -N OMEM H HI H " HN

C3gH44FIN6 0s:858.71 N 1N

a N0 18 C 2sH,,F FINO 4:515.55 NN

1 I(Ill)-SPIAd - MEM-2-Larotrectinib 8F-MEM-2- Larotrectinib

F F

OMEM N-N OH N'N 18F 18

IN HN6 18NN N 18F N N_ N6

0 0 18 1 C 2,H 30F FIN104 :515.55 C 2,H 24F aFIN10 2:427.45 1 18 F- MEM-2-Larotrectinib F- 2-Larotrectinib

[00487] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-MEM-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-MEM-2-Larotrectinib

detected that there was the labeled product [ 18F]-MEM-2-Larotrectinib.

[00488] Magnesium bromide (0.0033mmol, 0.6mg) was dosed in the reaction

mixture; the mixture was then heated to 155 °C to react for 10 minutes; the reaction

rate of 96.7% or above, the radiochemical purity of 96.3% and the chemical purity of

greater than 96.2%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was

37.29% relative to that in the V-shaped reaction flask (the radioactivity of 1[8 F] Et4NOMs

is usually 1.35mCi), and the specific activity (23.14Ci/ mol) was obtained from the final

preparation).

[00489] Example 127

[00490] Similar to the method of Example 126, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-MEM-5-Larotrectinib. The product had the deprotection rate of 97% and the

radiochemical purity of greater than 96% and chemical purity of greater than 96%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 37.25% relative to that in the

specific activity (23.21Ci/ mol) was obtained from the final preparation).

[00491] Example 128

[00492] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursor I(III)-SPIAd-THP-2-Larotrectinib.

F F

O N'N O N'N 0 H~I~18F0 HN N N 0 N N6N H 0 0 18 H C39HMFIN 6 0 7Si:854.72 C 26H30F FINS0 3:511.56 1 I(Ill)-SPIAd -THP-2-Larotrectinib 'F-THP- 2-Larotrectinib

F F

0O0N.-N OH N...N 18F N F NN

O O 1 1 C26 H 30F aFIN 603 :511.56 C 23H24F SFIN1O2 :427.45 18 18 F-THP-2- Larotrectinib F- 2-Larotrectinib

[00493] According to the preparation process of labeled product 1[ 8 F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-THP-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-THP-2-Larotrectinib

detected that there was the labeled product [ 18F]-THP-2-Larotrectinib.

[00494] P-toluenesulfonic acid (0.0033mmol, 0.56mg) was dosed in the reaction

mixture; the mixture was then heated to 155 °C to react for 5 minutes; the reaction

rate of 97.5% or above, the radiochemical purity of 98.2% and the chemical purity of

greater than 96.4%. The uncorrected radiochemical yield of [ 18F]-Larotrectinib was

41.32% relative to that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs

is usually 1.35mCi), and the specific activity (27.04Ci/ mol) was obtained from the final

preparation).

[00495] Example 129

[00496] Similar to the method of Example 128, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-THP-5-Larotrectinib. The product had the deprotection rate of 97% and the

radiochemical purity of greater than 99% and chemical purity of greater than 97%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 41.55% relative to that in the

specific activity (27.81Ci/ mol) was obtained from the final preparation).

[00497] Example 130

[00498] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-EE-2-Larotrectinib.

F F o0 0

18 H O1NN ON F NN N 1 N H, 0

18 H C 38H 44F1N60 7:842.71 C 25H 30F FIN 8O 3:499.55 1 |(Ill)-SPIAd -EE-2-Larotrectinib aF-EE- 2-Larotrectinib

F F

0S N -N O-0 \( NN OH

1F N N6S N N6

0 O 1 18 C 2sH 30F sFIN 603:499.55 C23 H24F FIN 602:427.45 18 18 F-EE-2-Larotrectinib F-2-Larotrectinib

[00499] According to the preparation process of labeled product [ 18F]-Larotrectinib,

anhydrous DMF-dissolved [1 8F]Et4NOMs solution (20mg/mL, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-EE-2-Larotrectinib (1.0mg) (the radioactivity of [ 18F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-EE-2-Larotrectinib

basically disappeared; radio-TLC tracked reaction mixture sampling (1-2uL) and it was detected that there was the labeled product 1[8 F]-EE-2-Larotrectinib.

[00500] P-toluenesulfonic acid (0.0033mmol, 0.56mg) was dosed in the reaction

rate of 99.8% or above, the radiochemical purity of 99% and the chemical purity of greater

than 99%. The uncorrected radiochemical yield of [ 18F]-2-Larotrectinib was 45.14%

usually 1.35mCi), and the specific activity (28.23Ci/ mol) was obtained from the final

preparation).

[00501] Example 131

[00502] Similar to the method of Example 130, labeled product

[ 18F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-EE-5-Larotrectinib. The product had the deprotection rate of 100% and the

radiochemical purity of greater than 99% and chemical purity of greater than 99%. The

uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 45.50% relative to that in the

specific activity (28.51Ci/ mol) was obtained from the final preparation).

[00503] Example 132

[00504] Labeled product [1 8F]-2-Larotrectinib was prepared from the labeled

precursorI(III)-SPIAd-Als-2-Larotrectinib.

F F -~0 0

07 N' N O- NN O

H- 11

" H~"

C3 7H 4eFIN 60sS:874.72 1 H C24H26 F sFIN 60 4S:531.57 1(II)-SPIAd-As-2-Larotrectiniib I(III)-SPIAd-As-2-Larotrectinib

F F

18 / -N O' 18 N OH F N §0 F NN

1 1 C24H 26F FIN 6 4S:531.57 C 2aH24 F FIN 60 2:427.45 1 |(ll)-SPIAd-As-2-Larotrectinib SF-2-Larotrectinib

[00505] According to the preparation process oflabeled product [ 18 F]-Larotrectinib,

anhydrous DMF-dissolved [ 18 F]Et4NOMs solution (20mg/L, 0.4mL) was prepared.

reaction flask containing the white solid labeled precursor

I(III)-SPIAd-Als-2-Larotrectinib (1.0mg) (the radioactivity of [ 18 F]Et4NOMs is usually

condition until TLC tracked that the labeled precursor I(III)-SPIAd-Als-2-Larotrectinib

detected that there was the labeled product [ 18 F]-Als-2-Larotrectinib.

[00506] Morpholine and 35% formic acid solution (1:1, 1.84mg) were dosed in the

reaction mixture, and acatalytic amount of palladium tetraphenylphosphorus was then

dosed; the mixture was then heated to 80°C toreact for 8minutes; thereaction solution was cooled in a 0 °C ice bath; 10% NaHCO3 aqueous solution was dropwise dosed until the solution was neutralized to neutral. The subsequent operation in this example was the same as that in Example 92. The product had the deprotection conversion rate of 99% or above, the radiochemical purity of 96.5% and the chemical purity of greater than 96.7%.

The uncorrected radiochemical yield of 1[ 8 F]-2-Larotrectinib was 41.54% relative to that

in the V-shaped reaction flask (the radioactivity of 1[ 8 F] Et4NOMs is usually 1.35mCi),

and the specific activity (26.72Ci/ mol) was obtained from the final preparation).

[00507] Example 133

[00508] Similar to the method of Example 132, labeled product

[ 1 8F]-5-Larotrectinib was prepared from the labeled precursor

I(III)-SPIAd-Als-5-Larotrectinib. The product had the deprotection rate of greater than

99% and the radiochemical purity of greater than 97% and chemical purity of greater than

97%. The uncorrected radiochemical yield of [ 18F]-5-Larotrectinib was 41.75% relative to

that in the V-shaped reaction flask (the radioactivity of [ 18F] Et4NOMs is usually 1.35mCi),

and the specific activity (26.71Ci/ mol) was obtained from the final preparation).

Claims

What is claimed is:

1. A [1 8 F]-labeled Larotrectinib compound, comprising a 1[ 8 F]-Larotrectinib compound

having the following structural formula and its analogs:

F 18 F

-NOH -NOH 1F (R) NF (R) N(R FHN N IHN N

1 0 2 O 18 18 F-2-Larotrectinib F-5-Larotrectinib

18F-2-Larotrectinib 18F-5-Larotrectinib

18 F

18F RN OH R1(2) N - N H

3 0 4 18 18 F-2-Larotrectinib analogues F-5-Larotrectinib analogues

18F-2-Larotrectinib analogues 18F-5-Larotrectinib analogues

2. A preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 1, wherein the method comprises the following steps:

step 1: hydroxyl protection: protection of the active hydroxyl group of an

iodo-Larotrectinib analog precursor (I-Larotrectinib);

step 2: preparation of a labeled precursor: carrying out iodine activation on the

iodo-Larotrectinib analogue precursor (I-Larotrectinib)with the activated active hydroxyl

group in step 1 to prepare a trifluoroacetic acid I-Larotrectinib analogue, and then causing

the trifluoroacetic acid I-Larotrectinib analogue to react with an adamantane-substituted

auxiliary acid to prepare a labeled precursor: hydroxy-protected spirocyclic hypervalent

I(III)-Larotrectinib analogue; and step 3: preparation of 1 8 F-labeled product: carrying out substitution reaction of the labeled precursor with a 18F radioactive source to prepare hydroxyl-protected

deprotection to obtain 1 8 F-Larotrectinib and a 18F-Larotrectinib analogue.

3. The preparation method of the 1[8 F]-labeled Larotrectinib compound according to

claim 2, wherein the I-Larotrectinib analogue precursor in step 1 has the following

structural formula:

R- NN OR 3

(R) N N HN N

in the above formula, RI and R2 each independently represent a phenyl substituent,

RI and R2 are each independently selected from group consisting of H, halogen, hydroxyl,

nitro, alkyl with 1 to 6 carbon atoms, alkenyl with 2 to 6 carbon atoms, alkynyl with 2 to 6

carbon atoms, cycloalkyl with 3 to 10 carbon atoms, aryl with 6 to 10 carbon atoms,

heterocycloalkyl with 4 to 10 carbon atoms, and heteroaryl with 6 to 10 carbon atoms; RI

and R2 each independently form a group with R3;

in the above formula, R3 specifically represents H;

the protection of the active hydroxyl functional group means that through esterification

or etherification, R3 is substituted with the following functional groups to protect the

active hydroxyl, and the substituted functional groups include trimethylsiloxane (TMS),

tert-butyldimethylsiloxane (TBDMS), triethylsiloxane (TES), tert-butyldiphenylsiloxane

(TBDPS), methyl ether (Me), benzyl ether (Bn), trityl ether (Tr), p-methoxytrityl ether

(MMT), dimethoxytrityl ether (DMT), tert-butyl ether ( tBu), methoxy methyl ether

(MOM), 2-methoxy ethoxy methyl ether (MEM), methylthio methyl ether (MTM),

benzyloxy methyl ether (BOM), p-methoxybenzyl ether (PMB), p-methoxybenzyloxy

methyl ether (PMBOM), 3,4-dimethoxybenzyl ether (DMB), tetrahydropyran ether (THP), methoxycarbonyl (Moc), ethoxycarbonyl (Eoc), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), acetyl (Ac), trifluoroacetyl (TfAc), chloroacetyl Base (CAc), dichloroacetyl (DCAc), benzoyl (Bz), pivaloyl (Pv), methanesulfonyl (Ms), benzylsulfonyl (Bs), allylsulfonyl (Bs), allyl

Alkyloxycarbonyl (Als), allyloxycarbonyl, alkanoyl with 1 to 16 carbon atoms,enoyl with

2 to 16 carbon atoms, alkynyl with 3 to 6 carbon atoms, cycloalkyl with 4 to 10 carbon

atoms, aroyl with 7 to 16 carbon atoms, and heterocycloalkanoyl with 4 to 10 carbon

atoms; alkanoyl is preferably acetyl; the aroyl is preferably benzoyl; the substituted

alkanoyl is pivaloyl or phenylacetyl.

4. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 2 or 3, wherein the I-Larotrectinib analogue precursor in step 1 has the following

structural formula:

FI

N OH N OH

N F F N

1-2- Larotractllb I5-L LaroiuinI or

5. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 4, wherein step 1 is to substitute the active hydroxyl in the I-Larotrectinib analog

precursor to achieve protection, and the substitution method comprises:

(1) benzoyl chloride esterification based on the following reaction formula:

F F 0

N _ _

OH N-NN NN OH0

1 C 28H 26FIN 603:640.46 C 21H22FIN60 2 : 536.35 1-2- Larotrectinib I - Bz-2-Larotrectinib

or,

F F

C21H 22 FIN 6 02 : 536.35 C 28H 26FIN 60 3:640.46 1 -5- Larotrectinib I - Bz-5-Larotrectinib

(2) pivaloyl chloride esterification based on the following reaction formula:

F F

N.N OH N' 0

C 21H 22FIN 60 2: 536.35 C 26H 30FINS0 3:620.47 I - 2-Larotrectinib I - Piv-2-Larotrectinib

or, II

N...N 0 N-N OH F OHF

C 21H22FIN 60 2: 536.35 C 26H30FINS0 3:620.47 I - 5-Larotrectinib I - Piv-5-Larotrectinib

(3) acetyl chloride esterification based on the following reaction formula:

F F

p NN OH N.--N 1 0

C 21H 22FIN 602: 536.35 C 2 3H24FIN0 3:578.39 I - 2-Larotrectinib I - Ac-2-Larotrectinib

or,

P NN OH N'-N OA

C21H22FIN602: 536.35 C23H24FIN603:578.39 I - 5-Larotrectinib I - Ac-5-Larotrectinib

(4) trifluoroacetic anhydride esterification based on the following reaction formula:

F F

ZPI NkN CF,

Nrf N

0 2 0

C 2 1 H2 2 F1N 6 02 : 536.35 C23H21F41N 603:632.36 I -2-Larotrectinib I -Tf-2-Larotrectinib or, 0

Z NN OH - N 0A F F -~FCF

0 2

C 21H 22F1N 60 2: 536.35 C 23H21F 4ING0 3:632.36 I-5-Larotrectinib I -Tf-5-Larotrectinib

(5) chioroacetyl chloride esterification based on the following reaction formula:

F F

-N OH / OOCCH 2CI

0 0

C2 1 H2 2 F1N 6 02 : 536.35 C23H23CIFIN 6O3: 612.83 1-2-Larotrectinib I -CAc-2-Laratrectlnlb

or,

NN OH 0 - N OOCCH 2CI F - F

N.CYF C21H 22FIN 60 2: 536.35 C23H23 CIFN 60 3:612.83 1- 5-Larotrectinib I -CAc-5-Larotrectinib

(6) dichloroacetyl chloride esterification based on the following reaction formula: F F

a'N - NN J OH 1I(N - NN H OOCCHC1 2

HN fNHN-I 0 0 C2 1 H 22 F1N 6 0 2 : 536.35 C23H 2 2cl2FIN603:647.27 1I-2- Larotrectinib I-DCAc -2- Larotrectlnib

or,

N'N OH N'N OOCCHC1 2

F NN N F O 0 0 C 21H 22 FIN 6 02 : 536.35 C 2zH 22 C 2 FIN 6O3:647.27 1 -5- Larotrectinib I-DCAc -5- Larotrectinib

(7) methyl chloroformate esterification based on the following reaction formula:

F F

N O N N O N N N N OO

C21 H 22FIN 6 0 2 : 536.35 C 23 H24 FING0 3:594.39 I --2-Larotrectinib I - Moc-2-Larotrectinib

or,

0 N- N OH N.-N OAO

N~ F NNF N6

1 -2

C21 H22 FIN 6 02 : 536.35 C 23 H24FIN 603:594.39 I --5-Larotrectinib I - Moc-5-Larotrectinib

(8) ethyl chloroformate esterification based on the following reaction formula:

F F

N N N 1 2 0

C 21H22FIN60 2: 536.35 C 24H 2 FIN60 4 :608.41 I- 2-Larotrectinib I -2- Eoc-Larotrectinib or,

F N P N H F N N

1 02 0

C21H22FIN6O2: 536.35 C24H2eFINSO4:608.41 I - 5-Larotrectinib I - Eoc-5-Larotrectinib

(9) Boc anhydride esterification based on the following reaction formula:

F F

C21H 22FIN 602 : 536.35 C26H30FIN6O4: 636.47 1 - 2-Larotrectinib I - Boc-2-Larotrectinib

(10) 2,2,2-trichloroethyl chloroformate (Troc) esterification based on the following

reaction formula:

F p N OH F NN O 0~l /N N~ N6 3

OH 0 ci 0 C 21 H2 2 FIN6 02 :536.35 3 FIN0 4:711.74 C 24H23C4 I -2-Larotrectinib I -Troc-2-Larotrectinib F F (11) trimethylsilyl ethoxycarbonyl chloride (Teoc) esterification based on the

followingreactionformula:

O OI

F F

N-N OH NN OTBS

C2 1 H22 FIN 6 02 :536.35 C 27 H 34 FIN6 4 Si:680.59 I -2-Larotrectinib I -Teoc-2-Larotrectinib

(12) tert-butyldimethylchlorosilane etherification based on the following reaction N N6N formula:

F OF

P NN OH N OTBS

C21 H 22 FIN 6 0 2 : 536.35 C27H36 FINS0 2Si:650.61

I - -LartrecinibI -TBS-2-Larotrectinib

(13) triisopropylchlorosilyl etherification based on the following reaction formula:

F F

N OTIPS -N OH N - N H N

ON 0 0

C21 H22 FIN 6 02 : 536.35 C30H42FIN602Si:692.69 I - 2-Larotrectinib I - TIPS-2-Larotrectinib

(14) tert-butyldiphenylchlorosilane etherification based on the following reaction

formula:

F F

I$~. ~N N OHN ON OBP N OTBDPS

N N

0 C 21H 22FIN 6 02: 536.35 C3 7H4 0FIN 602SI:774.75 I- 2-Larotrectinib I - TBDPS-2-Larotrectinib

(15) P-methoxybenzyl etherification (PMB-(p-Methoxybenzyl)ether) based on the

following reaction formula:

F F OMe

N OH N O

N NN; IN-I N~ KZI N6 NN J$

C21H22FIN 602 : 536.35 C2 9H 30FING0 3:656.50 I - 2-Larotrectinib I - PMB-2-Larotrectinib

(16) P-methoxybenzyl etherification (PMB-(p-Methoxybenzyl)ether) based on the

following reaction formula:

F F

aNJ -N OH N'N OMOM

NY H N HN 1 If20 0

C 21H 22FIN60 2:536.35 C 23H 2 8FING0 3:580.40 I - 2-Larotrectinib I - MOM-2-Larotrectinib

(17) methoxyethoxymethyl chloride etherification based on the following reaction

formula:

F F

N OH 0N...N OMEM 'ax,.".N

N N

1 2 o

C2 1 H2 2 FIN 6 0 2 : 536.35 C2 5H30FIN 604:624.46 1-2- Larotrectinib I - MEM-2-Larotrectinib

(18) 2-(trimethylsilyl)ethoxymethyl chloride etherification based on the following

reaction formula:

F F

O ..N-N OH N..--N 0

N 0 H N 0 N6 0

O~

(19) tetrahydropyran etherification based on the following reaction formula:

F F

/N.-N OI N OH

N N6

1 i 2 0

C 21H 22FIN 60 2: 536.35 C 26H 30FIN03:620.47 I - 2-Larotrectinib I -THP-2-Larotrectinib

(20) 1-ethoxyethanol acetate (EE) etherification based on the following reaction

formula:

F F 0

N OH N--N 0

N N N N6 HNO O 0 0

C 21H 22FIN 60 2: 536.35 C 25 H 30FIN 603 :608.46 I - 2-Larotrectinib I -EE-2-Larotrectinib

(21) allyl sulfonyl chloride etherification based on the following reaction formula:

F F 0

NN O' N-N OH

N NN ON-(

C 21 H 22 FIN6 0 2 : 536.35 C24H26 FINGO 4S:640.47 I - 2-Larotrectinib I - Als-2-Larotrectinib

(22) P-methoxyphenol etherification (PMP-(p-Methoxyphenyl)ether) based on the

following reaction formula:

F F OMe

OH N.N 0 NQN

N I N INN

1- 0

C 21H22FIN 602 : 536.35 C28 H2 8FING0 3:642.47 I - 2-Larotrectinib I -PMP-2-Larotrectinib

6. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 2, wherein the preparation of the trifluoroacetic acid iodo-Larotrectinib analogue in

step 2 is to cause the iodo-Larotrectinib analog precursor (I-Larotrectinib) with protected

active hydroxyl prepared in step 1 to react with trifluoroacetic acid or trifluoroacetic

anhydride, an organic solvent and an oxidant so that I is activated by trifluoroacetate;

the organic solvent comprises one or more of chloroform, dichloromethane,

acetonitrile, acetone, and tert-butanol peroxide;

the oxidant comprises a urea-hydrogen peroxide complex, Oxone,

2,2,6,6-tetramethylpiperidine-oxideormCPBA.

7. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 2, wherein the adamantane-substituted auxiliary acid in step 2 is SPIAd, and the

conditions for the reaction of SPIAd with the trifluoroacetic acid iodo-Larotrectinib analog

are as follows: SPIAd is mixed with one or more of sodium carbonate solution, MeCN,

and the mixed solution reacts with the trifluoroacetic acid iodo-Larotrectinib analogue at o0 C while continuously stirring for 1 hour to 10 hours, thus obtaining the labeled precursor.

8. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 2, wherein the preparation of the labeled precursor in step 2 specifically follows the

following methods:

(1) preparation of labeled precursor I(III)-SPIAd-Bz-2-Larotrectinib or

I(III)-SPIAd-Bz-5-Larotrectinib based on the following reaction formula:

F F

/-N OOCCsH, N'- N OOCCH,

F "I~ N(a NHN61 N N6 FCCOO H N,

0 0 C 28H 26FIN0 3:640.46 C32 H 28F 71N0 7:868.50

I - Bz-2-Larotrectinib I -TfAc- Bz-2-Larotrectinib and F F

OOCC6 H, 0 N'N OOCCH, F3 CCOO> N'N F3CCOO NH H N

O '"H N

Ca2H 2WFylN 60 7:868.50 H C41H 42FIN0 7:876.72 I-TfAc- Bz-2-Larotrectinib |(Ill)-SPIAd - Bz-2-Larotrectinib

(2) preparation of labeled precursor I(III)-SPIAd-Piv-2-Larotrectinib or

I(III)-SPIAd-Piv-5-Larotrectinib based on the following reaction formula:

F F 0 0 / -- N/ I ~~ F 3CCOO-.

FaCOOO N F3CCOO N 0 0 C 26H 30 FIN60 3:620.47 C 30 H-30 F 7 1N6 07 :846.50 I - Piv-2-Larotrectinib I- Piv-TfAc-2-Larotrectinib

F F CHcN-(y:86.5 000 CaH4F N8 y: -- 4 0

F 3CCOO N- H ~ N H3CO' N6 < HN--,

C30H-30F 71N 607: 846.50 H C39H"FINGO 7:854.72 I - Piv-TfAc-2-L-arotrectlnlb I(111-SPIAd -Pl-2-Larotrectinib

(3) preparation of labeled precursor I(III)-SPIAd-Ac-2-Larotrectinib or

I(III)-SPIAd-Ac-5-Larotrectinib based on the following reaction formula:

F F 0 0 JO r:- -N OA NN 0A H - O~H 3 F 3CCOO- 1 -J H N N6F 3CCOO N N~y HN N, HN--

, 0 0 C 23 H24FIN6 O3 :578.39 C 27H 24 F 71N0 7:804.42 I -Ac-2-Larotrectinib I -Ac-TfAc-2-Larotrectinib

F F 0 0

3 CO N ~N KZ OA _____N-_NO

F3 CN-- CH, N CH 3 VJCO- HN

C27H 24F 7 N 607: 804.42 H C36H 38F1N0 7: 812.64 1- Ac-TfAc-2-Larotrectinib I(11)-SPIAd -Ac -2- Larotrectinib

(4) preparation of labeled precursor I(III)-SPIAd-Tf-2-Larotrectinib or

I(III)-SPIAd-Tf-5-Larotrectinib based on the following reaction formula:

IF IF 0 -. 0 \/ 1 N- A0\ 51NN O F - ~ ~ CF3 F 3 CCOO... F (/X~~N N ~ F 3CCOO N HN NW HN-( 0 0 C 23H 2lF 41N03 :632.36 C 27H 23 CIFN0 7 :838.86 1-Tt-2-Larotrectinib I -Tf-TfAc-2-Larotrectinib

F F 0 0

-N OA0 N-N O ~ F3 CCOO- 1 F3CCOO 1 KIiN '--

HN- 3 H INI N NII C27H 21FI 0 1N07:858.39 H C 36H35F 4 N0 7: 866.61 1- Tf-TfAc-2-Larotrectinib I(111-SPIAd -Tf -2-Larotrectinib

(5) preparation of labeled precursor I(III)-SPIAd-CAc-2-Larotrectinib or

I(III)-SPIAd-CAc-5-Larotrectinib based on the following reaction formula:

F F

/ - N ~ OOCCHCI / N ~ OOCCHCI - \ F3CCOO-I

KII N HN N 3IIJN O0~ 0 0 C 23H 23C1F1N0 3 :612.83 C 27H 23CIFIN 607: 838.86 1-CAc-2-L-arotrectinib I -CAc-TfAc-2-L-arotrecti nib

F F

F3CCOO-. / 5;1 N-N OOCCH 2CI H 0 /- OOCHC 1

F~COO N HN- H, XO 00iJ

C27H 23CIFINGO7:838.86 H C36H 37CIFN0 7 : 847.08 1- CAc-TfAc-2-L-arotrectinib I(11)-SPIAd -CAc -2-Larotrectinib

(6) preparation of labeled precursor I(III)-SPIAd-DCAc-2-Larotrectinib or

I(III)-SPIAd-DCAc-5-Larotrectinib based on the following reaction formula:

F F

-~ N-N\ OOCCHC1 2 / -' N N ~ OOCCHC1 2 F 3CCOO-j N3 - F 3 CCOO N /N- N~ HN rII§ C23 H2 2 C12 FIN 6 0 3 : 647.27 C 27 H 22 C12 F 7IN 60 7 : 873.30 1- DCAc-2-Larotrectinib I - DCAc-TfAc-2-Larotrectinib

F F

F ~C- / 5-1 N - OOCCHC12 H -NN OOCCHC 2 1 F3CCOO 0N N6 N6I

C 27H22C1 2 F7IN60 7:873.30 C36H 36CI 2FIN 607: 881.52 1- DCAc-TfAc-2-L-arotrectinib H I(111-SPIAd -DCAc-2-Larotrectinib

(7) preparation of labeled precursor I(III)-SPIAd-Moc-2-Larotrectinib or

I(III)-SPIAd-Moc-5-Larotrectinib based on the following reaction formula:

OF 0F0

0 -0

- ~ j" ~F3CCOO- 1 N HN F 3CCOON

0 0 C 23 H24FIN 6O 4:594.39 C 27H 4 0F 7 N 608 :842.66 I -Moc-2-Larotrectinib I - Moc-TfAc-2-L-arotrectinib

F F 0 0

F3CCOO-I1 -J 1 F3CCOO' NA

C27H4OF 7IN 6O 8: 842.66 H C36H 38F1N0: 828.64 1-Moc-TfAc-2-Larotrectinib 1(111)SPIAd -Mc -2-Larotrectinib

(8) preparation of labeled precursor I(III)-SPIAd-Eoc-2-Larotrectinib or

I(III)-SPIAd-Eoc-5-Larotrectinib based on the following reaction formula:

F F 00

K§K N ~~ N H NrN F3CCOO-I F3CCOO KIIIIYN ~ N

0 0 C24H 26FING0 4:608.41 C 28 H 26F 7 1N0,: 834.44 1- Eoc-2-Larotrectinib I -Eoc-TfAc-2-Larotrectinib

F F 0 0

/ OA H 0 N- N 0 F3CCOO. 1 F3CCOO' NNN N6N

C28H 26F 7 N 608:834.44 H C37H40 FINSOS: 842.66 1- Eoc-TfAc-2-Larotrectinib I(11)-PIAd -Ec -2-Larotrectinib

(9) preparation of labeled precursor I(III)-SPIAd-Boc-2-Larotrectinib or

I(III)-SPIAd-Boc-5-Larotrectinib based on the following reaction formula:

N- N OBoc 0 N - OBoc

Cy HJN, H". 0 0

C2 6H 30 F1N04 : 636.47 H C 3 gH"FIN6 Oa: 870.72 1- Boc-2-Larotrectinib I(111-SPIAd -Boc-2-Larotrectinib

(10) preparation of labeled precursor I(III)-SPIAd-Troc-2-Larotrectinib or

I(III)-SPIAd-Troc-2-Larotrectinib based on the following reaction formula:

F F OCCCOi

~ 3CCOO. N H N6~"FCO HN..-(N..

0 0 C2 4 H 2 3 C1 3 FIN 6 0 4 :711.74 C 28 H23 C13 F7 1N 6 0 8 :937.77 I -Troc-2-Larotrectinib I -Troc-TfAc-2-Larotroctinib

F 0 CC 3 F 3 CCOO3 0 5- NII F 3CCOd- N. . [). , 6 ~j

C28 H2C1 3 F7 IN 6O 8 :937.77 H C37 H 37 C13 FIN 60 8 :945.99 I -Troc-TfAc-2-Larotrectlnlb I(111-SPIAdTroc-2-Larotrectinib

(11) preparation of labeled precursor I(III)-SPIAd-Teoc-2-Larotrectinib or

I(III)-SPIAd-Teoc-5-Larotrectinib based on the following reaction formula:

F 0F0

5; - N 0 k /O/ 5; N-N -. F 3 CCOO- 1 N H N6~1~ F3CCOO K111 SiSi- N H N..§

0 0 C 27 H34 F1N 6 0 4 Si:680.59 C31 H 34 F7 IN 6 0O8 Si:906.63 I -Teoc-2-Larotrectinib I -Teoc-TfAc-2-Larotrectinib

F F

KlIi 1 F3 CCOO HN fN6 0' \ HN.-.N

C3 1 H3 6 F7 1N 6 07 Si:892.64 H C 40H 4SFTN60 8 Si:914.85 I -Teoc-TfAc-2-Larotrectinib I(111-SPIAd -Teoc-2-Larotrectinib

(12) preparation of labeled precursor I(III)-SPIAd-TBS-2-Larotrectinib or

I(III)-SPIAd-TBS-5-Larotrectinib based on the following reaction formula:

F F

~'~- N OTBS /x N ~ OTBS F3CrCOO- 1 \

N N6- - F 3CCOO / N HN-W N N

C2 7H 36FIN 602S1:650.61 C~1 H36F7IN 606 Si:876.64 1-TBS-2-Larotrectinib I -TBS-TfAc-2-Larotrectinib

F F

F CO-~'~/~ N-N OTBS 0 XNN T FCCOO'3 I _ H CJ NS

N HN- H,( N

C 31H 36F 71N 6 6Si: 876.64 H C4 0H 50FING06 8:884.86 1 -TBS-TfAc-2-Larotrectnlb I(111-SPIAd -TBS -2-Larotrectinib

or,

N ~ OTBS 0 ~ NN ~ OTBS N ,N6NN H --

C 27H 36FING0 2S1:650.61 H C40 H5 0FINB0 6S: 884.86 1-TBS-2-Larotrectinib I(111-SPIAd -TBS -2-Larotrectinib

(13) preparation of labeled precursor I(III)-SPIAd-TBDPS-2-Larotrectinib or

I(III)-SPIAd-TBDPS-5-Larotrectinib based on the following reaction formula:

F F

N -N OTBDPS ' -NCO OTBDPS -~~~ F 3CCOO N H

C 37H 40FINS0 2 Si:774.75 C4 1H 40F 71N 6Si: 1000.79

1-TBDPS-2-Larotrectinib I -TBDPS-TfAc-2-Larotrectinib

F F

N OTBDPS 0 N OTBDPS F3CCOO-I HI F3CCOO N '2 N6 IIIN 0 H H

C4jH4OF 7IN 6 O6Si:1000.79 H Cs50 H4FIN6O 6Si: 100901 I -TBDPS-TfAc-2-Larotrectlnlb I(111-SPIAd -TBDPS -2-Larotrectinib

(14) preparation of labeled precursor I(III)-SPIAd-TIPS-2-Larotrectinib or

I(III)-SPIAd-TIPS-5-Larotrectinib based on the following reaction formula:

F F

/ ~ ~ CO ~ N 5 OTIPS 1a N- 3 '/~N - OTIPS

CN N F 3 CCOO 1 N N N~

0 0 C3 0H-42FIN 602S1:692.69 C 34H-42 F 7 1N6 06S: 918.72 1- TIPS-2-Larotrectinib I -TIPS-TfAc-2-Larotrectinib F F

/~ N ~ OTIPS -0 N OTIPS 3CCO N N FCO N N HN-, N,"C N6 0i 00

C 34H 42F 71N 6 06 S: 918.72 H C43H-56 FIN 6O6 Si:926.94 1- TIPS-TfAc-2-Larotrectinib I(11)-SPIAd -TIPS -2-Larotrectinib

(15) preparation of labeled precursor I(III)-SPIAd-PMB-2-Larotrectinib or

I(III)-SPIAd-PMB-5-Larotrectinib based on the following reaction formula:

F OMe F p ~ ~ N 0 N\I \ F 3CCOO- 10

K NbN6 F3CCOO1 N;i HN

C 33 H30 F7 1N6 0 7 :882.53 C29H 30FIN 603:656.50 I -PMB-TfAc-2-Larotrectinib I -PMB-2-Larotrectinib

F3 CCOO- 1 \H 4 F3CCOO C H

C3 3H 3oF 7 1N 60 7 :882.53 H C4 2H44FIN 6O 7 :890.75 I -PMB-TfAc-2-Larotrectinib 1(111-SPAd -PMB-2-Lamotnctinib

(16) preparation of labeled precursor I(III)-SPIAd-MOM-2-Larotrectinib or

I(III)-SPIAd-MOM-5-Larotrectinib based on the following reaction formula:

F F

N CO O -N~ OMOM

0I'IIN HN 0~I$- F3CCOO KN N HN- -;III C 23 H26 F1N 6 0 3 :580.40 C 27 H 26 F7 1N6 07 :806.43 I -MOM-2-Larotrectiriib I - MOMTAc-2-1-arotrectinib

F F

N/ OMaN 0 - N\ OMOM F3CCOO- 1 7H

( F3CCuu N N6H~OK \ HN--(

C 27 H26 F 7]N 6 07 :806.43 HC36H 4 OFIN 6O 7 :814.65 I -MOM-TfAc-2-L-arotrectlnlb I(11)-SPIAd -MOM-2-Laratrectinib

(17) preparation of labeled precursor I(III)-SPIAd-MEM-2-Larotrectinib or

I(III)-SPIAd-MEM-5-Larotrectinib based on the following reaction formula:

F F

~~ F3CCOO - ME

N N F3 CCOO' 7J N

C25 H 30 F1N 6 0 4 :624.46 C29 H3 0 F71 N 6 O8 :850.49 I -MEM-2-Larotrectinib I -MEM-TfAc-2-L-arotrectinib

F F

- /N OMEM 0 0/N NCO OMEM F3C'- HI

F 3CCOO IJN N N6 H N N:~1 C 29 H3 0 F7 1N 6 08 :850.49 H C 38 HluFIN 6O 8 :858.71 I -MEM-TfAc-2-Larotrectinib I(111-SPIAd - MEM-2-Larotrectinib

(18) preparation of labeled precursor I(III)-SPIAd-SEM-2-Larotrectinib or

I(III)-SPIAd-SEM-2-Larotrectinib based on the following reaction formula:

F F

/ - -F N ~ 0 " f - N ~ OSEM

3 CCOO- 1 S.\ N N6F 3CCOO N N-, HN-r N iN

0 0

C 27 H 36 FIN 6 0 3 Si:666.61 C 31 H 36 F7 1N 6 0 7 Si892.64 I -SEM-2-Larotrectinib I -SEM -TfAc-2-L-arotrecti nib

F F

F3 CO-N~'~ OSEM H N /N OSEM 3CCOO-1 ~ H'I O C N

C3 jH36F 71N60 7 Si:892.64 H C40 H50 FIN 60 7 Si:900.86 I -SEM-TfAc-2-Larotrectinib I(111-SPIAd -SEM-2-Larotrectinib

(19) preparation of labeled precursor I(III)-SPIAd-THP-2-Larotrectinib or

I(III)-SPIAd-THP-5-Larotrectinib based on the following reaction formula:

F OxF

I ~~ -Nt J F 3CCOO> /IN HN-I 0NH

0 0 C27 H 30 FIN 6 O 3 Si:620.47 C3 9 H44FPN6 O 7 Si:854.72 I -THP-2-Larotrectinib I -TfAc- THP-2-Larotrectinib

0 0

F3 CCOO-I F 3 CCOO0- KililN H C39 H44FLN 6 O 7 Si:854.72 C39H4FJIN6O 7Si:854.72 I-TfAc- THP-2-Larotrectinib I(I1)SPIAd -THP-2-Larotrectlnlb

(20) preparation of labeled precursor I(III)-SPIAd-EE-2-Larotrectinib or

I(III)-SPIAd-EE-5-Larotrectinib based on the following reaction formula:

F F

_________ F 3 CCOO-I

N N6F 3CCOO /~ HN NW HN-, 0 0

C 26H 3 0FING0 3:608.46 C2 9 H30 F1N 6 0 7 :834.49 I -EE-2-Larotrectinib I -TfAc- EE-2-Larotrectinib

F F

0 \ 0 / N H 0 I -O 0/ F3CCOO- 1 00 \

F 3CCOO />I N6jiii j

0 H H C 29H 30 FflN 607 :834.49 HC 3 sH44 F1N 6 07 :842.71 I -TfAc- EE-2-Larotrectinib I(11)-SPIAd -EE-2-Larotrectinib

(21) preparation of labeled precursor I(III)-SPIAd-Als-2-Larotrectinib or

I(III)-SPIAd-Als-5-Larotrectinib based on the following reaction formula:

F F

10 " N F3 CCOO N N N6 F 3CCOO h \

(\jHN N (j HN

C2 5H30 FIN0 4S:656.52 C 29H 3 0F 7IN 60 6S:882.55 I - As-2-Larotrectinib I - As-TfAc-2-Larotrectinib

F F 0 N-0 XN- N'N O I NN O-I F3CCOO-I O H F3CCOd' 'ilJ N HN6 _-N 1Z

C 29H 3oF 71N 6 0 6S:882.55 H C3 7H40FIN6O 8S:874.72 I -Als-TfAc-2-Larotrectinib |(Ill)-SPIAd -Als-2-Larotrectinib

(22) preparation of labeled precursor I(III)-SPIAd-PMP-2-Larotrectinib or

I(III)-SPIAd-PMP-5-Larotrectinib based on the following reaction formula:

F OMe F

al: - _. OMe N-N~~~

N N F 3CCOO N Hi HN F3CCO N6 HN _ N

0 0 C 28H 28FIN 603:642.47 C32 H28F7 1N 60 7 :868.50 1 - PMP-2-Larotrectinib I - PMP-TfAc-2-Larotrectinib

F F

N-NI OMe N OMe F 3CCOO-.H F 3CCOO N HN

0 H0 C32H 28 F7 1N 6 07 :868.50 H C41H 42FIN 6 0 7 :876.72 I - PMP-TfAc-2-Larotrectinib I(III)-SPIAd -PMP-2-Larotrectinib

9. The preparation method of the 1[ 8 F]-labeled Larotrectinib compound according to

claim 2, wherein the 1 8 F radioactive source in step 3 is obtained by the following method:

(1) preparation of [1 8 F] fluoride target water: producing [1 8 F] fluoride target water by

1 80(p,n) 1 F8 nuclear reaction, and delivering the [ 1 8F] fluoride target water to a sterile

lead-protected hot cell of 1 8 0-enriched water by nitrogen pressure;

(2) enrichment of [1 8F] fluoride by QMA anion exchange solid phase extraction

cartridge (QMA): causing an aliquot of the target water containing an appropriate amount

of [1 8F] fluoride to slowly pass through the anion exchange solid phase extraction

cartridge (QMA) under a nitrogen flow, enriching the [1 8F] fluoride on the QMA anion

exchange solid phase extraction cartridge (QMA), and removing 180 and other impurities

by separation and elution, thus obtaining [ 18F] fluorine source for the [ 18F] fluoride QMA

anion exchange solid phase extraction cartridge (QMA);

(3) eluting the 18

[ F] fluoride enriched on the QMA anion exchange solid phase

extraction cartridge (QMA) to obtain a quaternary ammonium salt or inorganic salt

solution of [18 F] fluoride; washing the [18 F] fluoride enriched on QMA anion exchange

solid phase extraction cartridge (QMA) with a washing solution and eluting the [ 18F]

fluoride into a V-shaped flask sealed with a Teflon-lined diaphragm, thus obtaining an

acetonitrile aqueous solution or an acetonitrile or methanol solution of the organic or

inorganic salt of [ 18F] fluoride; and

(4) preparation of a dry quaternary ammonium salt or inorganic salt of [ 18F] fluoride:

heating the V-shaped flask containing the acetonitrile aqueous solution or acetonitrile or

methanol or ethanol solution of the organic or inorganic salt of [ 18F] fluoride and sealed

with a Teflon-lined diaphragm to 95 'C to 110 'C, and at the same time drying nitrogen

gas by passing through a P205-DrieriteTM column to blow the V-shaped flask, and then

discharging the exhaust gas through the vented flask; when no liquid is seen in the flask,

taking the flask out of the hot bath, dosing anhydrous acetonitrile (1 mL) in the flask, and

heating the flask again until the flask is dry; repeating this step another three times; then,

cooling the flask at room temperature under nitrogen flow, thus obtaining a dry organic or

inorganic salt of [ 18F] fluoride.

10. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 9, wherein the anion exchange solid phase extraction cartridge (QMA) in step (2) is

pre-activated by being washed with NaHCO 3(aq) and water;

the washing solution in step (3) is obtained by dissolving an organic or inorganic base

in acetonitrile and water (v/v: 7:3) or acetonitrile or methanol or ethanol, and the organic

or inorganic base comprises tetraethylammonium bicarbonate;

the organic or inorganic salt of [ 18F] fluoride in step (4) comprises

[ 18F]KF/K 2 CO 3 /K 2 .2.2, [ 18F]KF/K2C204/K2.2.2, [ 18F]KF/KOTf, [ 18F]Et4NF, [ 18F]Et4NHCO3,

[ 18F]Et4NOMs, and [ 18F]Et4NOTf, and its radioactive [ 18F] fluoride recovery rate varied depending on the elution process used.

11. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 9, wherein the substitution reaction between the labeled precursor and the 18F

radioactive source in Step 3 is implemeted by: dosing a solvent in the V-shaped flask

containing the dry organic or inorganic salt of [ 18F] fluoride to re-dissolve the dry organic

or inorganic salt, and then dosing the labeled precursor to take reaction, thus obtaining a

crude reaction solution of undeprotected labeled product 18

[ F]-Larotrectinib.

12. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 9, wherein deprotection in step 3 is implemented by the following method: dosing or

solution of labeled product [1 8F]-Larotrectinib.

13. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 12, wherein the crude reaction solution of labeled product [ 18F]-Larotrectinib is

of 20% Tween 80 ethanol solution is a labeled product [ 18F]-2-Larotrectinib injection.

14. The preparation method of the [ 18F]-labeled Larotrectinib compound according to

claim 2, wherein the preparation of [ 18F]-labeled Larotrectinib from the labeled precursor

in step 3 comprises the following methods:

(1) labeled product [ 18F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Bz-2-Larotrectinib based on the following reaction formula:

F F

0 N'N OOCCHu N.N OOCC8 H, H O F

H H 0 0

H C4 1H42FIN 6 07:876.72 C2 8H 26F'SFN 60: 531.55 I(Ill)-SPIAd - Bz-2-Larotrectinib OF -Bz-2-Larotrectinib

F F

F N OCC6Hs NN OH 18 N:FN FN N HN N 0 0 8 C2 8H 26F' FN 6 0: 531.55 C2 1H 2 2F'sFN60 2 : 427.45 18 18 F -Bz-2-Larotrectinib F -2- Larotrectinib (2) labeled product [ 8F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-TBS-2-Larotrectinib based on the following reaction formula:

F F

0 N'N OTBS N.N OTBS H _S

H 0 N F

18 H C 40HS 0FIN 6 06Si:884.86 C 27 H36F FN6 O2 Si:541.71 1 I(Ill)-SPIAd -TBS -2-Larotrectinib F -TBS-2- Larotrectinib

F F

18 F dN.N HNN OTBS 18 F

N NNO

8 C2 7H,,F'SFN 6O2 Si:541.71 C21 H22F1 FNI02: 427.45 18 18F -TBS-2- Larotrectinib F - 2-Larotrectinib

(3) labeled product [ 18F]2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-SEM-2-Larotrectinib based on the following reaction formula:

FF

0 - OSEM N OSEM I FN H

18 H C 40 H50 F1N 6 07 Si:900.86 C 27 H36 F FN 6 03 Si:557.71 F (11)-SPIAd -SEM-2-Larotrectlnlb 18F -SEM- 2-Larotrectinib

18F N'N P, J OSEM :P~ ' N -N OH

a N A ?,r § N K 5 C 21 2 ,F'SFN 6 0 2 :427.450 C 27 1 36 F'SFN6 0 3 Si:557.71 18 18 F -SEM- 2-Larotrectinib F -2- Larotrectinib

(4) labeled product [F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-TBDPS-2-Larotrectinib based on the following reaction formula:

~~;/ - N OTBDPS / Ni OBP H 19F<K

"H0 0 19 H C 5 0HrAFIN 6O 6Si: 1009.01 C3 7HF FIN1O2Si:666.85 1 I(111-SPIAd -TBDPS -2- Larotrectinib 9F-TBDPS-2-Larotrectinib

F F

/-N OTIBDPS N OH 19F 19F

9 19 C3 7H 40F' FI10 2S1:666.85 C 21H-22 F FIN 602:428.44 19 19 F -TBDPS-2-Larotrecti nib F -2- Larotrectinib

(5) labeled product [1 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-TIPS-2-Larotrectinib based on the following reaction formula: F IF

0iN OTIPS N TIPS

HH1)SId-IS2Lrorcii 19FIP--arteci 0 143

F F

N-/ OTIPS N9 OH

9 9 19F 2F 1 6 0S:547 19F 2F 1N0:48

19 'OF-TIPS-2-Larotreetinib F -2-Larotrectinib

(6) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Teoc-2-Larotrectinib based on the following reaction formula:

XF 0F0 0 0

0 0 1 H C 40H 4 SFIN 60 8 Si:914.85 C 27 H34F FN6 O4 SI:571.69 18 I(111-SPIAd -Teoc-2-Larotrectinib F -Teoc- 2-Larotrectinib

FF O

0

F F8

N6 HHN -i HNI 1F Q N K 0 H 2H4'8N0 C 35. 9H~I 67847 C2, 2H0FN 60: 1.56 18F -T1o- P - Piv-2-Larotreci1iF -- 2Larotrectinib

F F 00

/N--kr 0N 0O

0

18 18 H 3 0F 603 :5115 1NCH 144 C2 H F 2 F 603:.45 I PIQ- Pd-Paoc iv-2 Laarctnb i-2-Larotrectinib

(8) labeled product [18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Ac-2-Larotrectinib based on the following reaction formula:

F F 0 0

O N'N OA5, N'N OAC3

18 HN-- IYN OH 3 C$H F NKJ H O

[I00 HN -H 0 O H C 36H 38FIN 607:812.64 8 C23H24F FIN 6O 3 :469.48 I(Ill)-SPIAd -Ac - 2-Larotrectinib 1OF - Ac-2-Larotrectinib

F F 0

-N O N ~ OH 18 18 F N H3 F N N I) N6 c N HN N---I O 0 C 23 H24 FIOFIN 60 3:469.48 C 2,H 24FiOFIN 602 :427.45 1 18 F - Ac-2-Larotrectinib F- 2-Larotrectinib

(9) labeled product [ 18F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Tf-2-Larotrectinib based on the following reaction formula:

F F 0 0

O N'N O C N'-N OA 18 NN CF 3 H NF 3

H 00 H C 36H 3OF 4 1N 607:866.61 18 C 23H 21F4 FIN 60 3:523.45 |(li)-SPAd -Tf-2-Larotractinib 18 F-Tf-2-Larotrectinib

F F 0

N-N O CF N -N OH 18 18 F N$N F3 F N YN HN6

0 0 18 1 C23 H21F 4 FIN 603:523.45 C2 3H 24F sFIN10 2 :427.45 1 1OF - Tf-2-Larotrectinib F- 2-Larotrectinib

(10) labeled product [ 18F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-CAc-2-Larotrectinib based on the following reaction formula:

F F 0~ 0

0N-N H ICH 2CI 18 FCHC /A 5'1 K$C OA2

H, ONN N H~

H 18 C 36H37CIF1N0 7: 847.08 C 23H23CIF FIN03 :503.93 I(11)-SPIAd -CAc -2-Larotrectinib 'OF -CAc-2-L-arotrectinib

F F

N0 18 F - \ J CHCi 18 F

18 C 23 H2 3CIF F1N 60 3:503.93 C 23H 2 4F'8FI1 6 2 :427.45 18 'OF-CAc-2-Larotrectinib F-2- Larotrectinib

(11) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-DCAc-2-Larotrectinib based on the following reaction formula:

F F

0 0 0

HHCHCI 2 18 CHC 2

H 18 C 36H-3 0C12F1N 60 7: 881.52 C 23H22C1 2F FIN 603:538.37 18 I(11)-SPIAd -DCAc -2-Larotrectinib F -DCAc-2-Larotrectinib F F 0

-N 0 -N OH 18 F - \ J CHCI 2 18 FJ

18 C 23 H2 2Cl 2FIOF1N 60 3:538.37 C 23H 24F F1NS0 2:427.45 18 18 F -DCAc-2-Larotrecti nib F-2- Larotrectinib

(12) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Moc-2-Larotrectinib based on the following reaction formula:

F F 0 00 0 / - N 0_ _ 1A x~ / 5; N--kO

H I 18CJF 0HN 0 0

H C 30H38FIN 608: 828.64 146 18 C23H-24F FINS0 4 :485.48 I(111-SPIAd -Moc-2- Larotrectinib 1F- Moc-2-Larotrectinib

F F 0 18 OH N 0 /0 N\ 18 IF NFN HN I

8 C 23H 2 4FIOF1Nr 604 :485.48 C2 3H 24F' FIN60 2:427.45 I- Moc-2-Larotrectinib 'OF- 2-Larotrectinib

(13) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Eoc-2-Larotrectinib based on the following reaction formula:

F F 0 0

HX I 5- Nj 18F0

H 0 0 H C37H 40FIN60 8 :842.66 18 C 24H 26F F1N 604 :499.51 I(111-SPIAd -Eoc-2- Larotrectinib 'F- Eoc-2-Larotrectinib

F F 0

NQ N 0 0 ~N 0H

8 C 24H 26 F' FIN6 04:499.51 8 2 :427.45 C2 3H 24F'SFIN ,0 18 'OF- Eoc-2-Laratrectinib F-2- Larotrectinib

(14) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Boc-2-Larotrectinib based on the following reaction formula:

F18 F H 0

18 H C39H 44FIN60 8 : 870.72 C 26H30F F1N04:527.56 18 I(11)-SPIAd -Boc-2- Larotrectinib F -Boc-2-Larotrectinib

F 0/ F

0 18 F 18NF

0N 1470 8 C 26 H3 0FIOF1N 604 :527.56 C2 3H 24FI FIN60 2:427.45

(15) labeled product [ 8 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Troc-2-Larotrectinib based on the following reaction formula:

F F

O /'N CC 3 NO

H H H 10F 18F N N 013

8 H C 37H37C13FIN60S:945.99 C 24H 23CI 3F FIN 6O 4:602.84 1 |(lll)-SPIAd -Troc-2-Larotrectinib F -Troc-2-Larotrectinib

F F 0

N-N 0 O N'N OH 18F 3F 8N N

0 0 18 1 C 24 H23Cl 3F FIN 604:602.84 C 23H 24F sFINS0 2:427.45 18 1 F- Troc-2-Larotrectinib 8F- 2-Larotrectinib

(16) labeled product [ 8F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Troc-2-Larotrectinib based on the following reaction formula:

F F

-~~ CC1 3 O N'-N O0 O C-l N'-N OA

H, O N8F N N6 CC13 ON-, 0NH H 0 "'Lo 18 H C37H 37Cl3FIN60 8:945.99 C 24H 23C1 3F FIN 604:602.84 |(III)-SPIAd-Troc-2-Larotrectinib 'F-Troc-2-Larotrectinib

F F

-N OH 18 18 F F N HN--, (CNN ~ N-& \J N

18 18 C 24H 23C1 3F FING04 :602.84 C 23H24 F FINI02 :427.45 18 18 F- Troc-2-Larotrectinib F- 2-Larotrectinib

(17) labeled product [ 18F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-PMB-2-Larotrectinib based on the following reaction formula:

\-x Ome -O OMe

H_ _ 18F

1 H C4 2H44F][N 6O,:890.75 C 2 9H 30 F "FIN 6 O3 :547.60 I(111-SPIAd -PMB-2-Larotrectinib I(11)-SPIAd -PMB-2-Larotrectlnlb

F F

[aOMe

18 N 0/ OH

18 18 C29 H30 F F1N 6 03 :547.60 C 2,H 2,F F10 2:427.45 18 I(111-SPIAd -PMB-2-Larotrectinib F- 2-Larotrectinib

(18) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-MOM-2-Larotrectinib based on the following reaction formula:

F F

0 N OMOM H0 - N , 5; OMOM 18 0 F

H,,. 0 ALHN-N ' N (2 N HN NJ 18 H C 36H40 F1 60 7:814.65 C 23H 26F FINS0 3:471.50 8 I(111-SPIAd - MOM-2-Larotrectinib I F-MOM- 2-Larotrectinib

F F

N OMOM -N OH

N ~

8 C 23H 2 6F' FI10 3:471.50 C 23H 24F'gF10 2:427.45 18 18 F- MOM-2-Larotrectinib F- 2-Larotrectinib

(19) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-MEM-2-Larotrectinib based on the following reaction formula:

F F

0 / OMEM 1N-N OE

H 0N 149 0a

8 H C 38H44 FIN6 Os:858.71 C 25H 30FI F1N 60 4:515.55

F F

N-N OMEM N' OH 18 F ~. 1pN N F N N;N6

0 0 C26 H 30FisFIN 6O4:515.55 C 23H 24FiOFIN O 6 2:427.45 1 18 F-MEM-2-Larotrectinib F-2-Larotrectinib

(20) labeled product [ 8F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-THP-2-Larotrectinib based on the following reaction formula:

F F

0 - -0 0 _1N 00

N' N6O ' N O O-N H H O N 0 1aF THN-, 0 H P 18 H C39H44FIN60 7 Si:854.72 C26H3 0F FINSO3:511.56 18 |(Ill)-SPIAd -THP-2-Larotrectinib F-THP- 2-Larotrectinib

F F

18 'OF - -N_( F N N6O 1N N6iN HN N

0 0 1 1 C 26 H 30F sFIN 603:511.56 C 23H 24F FIN 6 02 :427.45 1 1 sF-THP-2- Larotrectinib sF- 2-Larotrectinib

(21) labeled product [ 18F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-EE-2-Larotrectinib based on the following reaction formula:

F F

/ X:;, ~N- 0 18 H 3yN HN6(I F N HN6N

18 H C38 HE 44 F1N6 0 7 :842.71 C28H30F FIN60 3 :499.55 18 I(111-SPIAd -EE-2-Larotrectinib F-EE- 2-Larotrectinib

F F

'~-/ N~ N~ OH _ _F 18F

8 18 C 2,H 3,F' FI10 3 :499.55 C2 3H24F FIN 60 2 :427.45 18 18 F-EE- 2-Larotrectinib F2- Larotrectinib

(22) labeled product [ 18 F]-2-Larotrectinib is prepared from the labeled precursor

I(III)-SPIAd-Als-2-Larotrectinib based on the following reaction formula:

F F 00

oIB

0 N N oil>

F Fo

00

18F

18F N ~ 0

N HN Ki i 0 0 8 18 C2 4H,,F' F11N60 4S:531.57 C 23H 24 F F1N 602:427.45 1 I(111-SPIAd -As-2-Laratrectinib $F-2- Larotrectinib